Macrocyclic compounds useful as pharmaceuticals

ABSTRACT

The present invention provides methods for the use of compounds having formula (I) in the treatment of various disorders including inflammatory or autoimmune disorders, and disorders involving malignancy or increased angiogenesis, wherein R 1 -R 11 , t, X, Y, Z, and n are as defined herein.

PRIORITY CLAIM

This Application is a divisional of U.S. application Ser. No.10/507,067, filed Nov. 10, 2004, which is a 35 U.S.C. §371 filing ofInternational Application Number PCT/US03/07377, filed Mar. 7, 2003,which claims priority to U.S. Provisional Patent Application Nos.60/362,883, filed Mar. 8, 2002, and 60/380,711, filed May 14, 2002, eachof which is incorporated herein by reference in its entirety. Thecontents of any patents, patent applications, and references citedthroughout this specification are hereby incorporated by reference intheir entireties.

BACKGROUND OF THE INVENTION

F152 (LL-Z1640-2) (1) is a zearalenone-like macrolide, isolated fromshake flask fermentation, crude extracts of which inhibited the ciliatedprotozoan Tetrahymena pyriformis (see, McGahren et al. J. Org. Chem.1978, 43, 2339). It was reported that initial biological studies usingthis natural product failed to yield any particularly interestingactivities.

After initial isolation and reporting of this compound, several othergroups explored the possibility of preparing additional derivativesand/or further exploring their biological activity. For example,scientists at Merck reported that F152 and certain isomers thereofinhibit the phosphorylating enzyme Map/Erk kinase (MEK) and thus areuseful for the treatment of certain cancers and other diseasescharacterized by the formation of neoangiogenesis (see, GB 323 845).Other groups have also reported derivatives of F152 having activity astyrosine kinase inhibitors, which are useful, for example, for thetreatment of cancer and inflammatory disorders (see, EP 606 044; WO00/38674; JP 8-40893; WO 96/13259; U.S. Pat. Nos. 5,728,726; 5,674,892;5,795,910). Each of these groups, however, was only able to obtain F152and derivatives thereof by fermentation techniques and by modificationsto the natural product, respectively, and thus were limited in thenumber and types of derivatives that could be prepared and evaluated forbiological activity. Additionally, although F152 and certain derivativesthereof have demonstrated potent in vitro activities, these compoundsare biologically unstable (for example, they are susceptible to enoneisomerization in mouse and human plasma), thereby limiting thedevelopment of these compounds as therapeutics for the treatment ofhumans or other animals.

Clearly, there remains a need to develop synthetic methodologies toaccess and examine the therapeutic effect of a variety of novelanalogues of F152, particularly those that are inaccessible by makingmodifications to the natural product. It would also be of particularinterest to develop novel compounds that exhibit a favorable therapeuticprofile in vivo (e.g., are safe and effective, while retaining stabilityin biological media).

SUMMARY OF THE INVENTION

As discussed above, there remains a need for the development of novelanalogues of F152 and the evaluation of their biological activity. Thepresent invention provides novel compounds of general formula (I),

and pharmaceutical compositions thereof, as described generally and insubclasses herein, which compounds are useful as inhibitors of NF-κBactivation, AP-1 activation and protein kinases (e.g., MEKK1, MEK1,VEGFr, PDGFr), exhibit antiangiogenic activity, and/or have ananti-anflammatory effect. Thus these compounds are useful, for example,for the treatment of various disorders including inflammatory orautoimmune disorders, and disorders involving malignancy or increasedangiogenesis. The inventive compounds also find use in the prevention ofrestenosis of blood vessels subject to traumas such as angioplasty andstenting.

DESCRIPTION OF CERTAIN PREFERRED EMBODIMENTS OF THE INVENTION

In recognition of the need to access and further explore the biologicalactivity of novel analogues of F152, and this class of macrocycles ingeneral, the present invention provides novel macrocyclic compounds, asdescribed in more detail herein, which demonstrate increased stabilityand are potent inhibitors of NF-κB activation, AP-1 activation andprotein kinases (for example, MEKK, MEK1, PDGFr, VEGFr). Based on thesemechanisms of action, the compounds inhibit the production of variouspro-inflammatory and/or immunologic cytokines such as TNFα, IL-1, IL-6,IL-8, IL-2 etc, and also inhibit the production of variouspro-inflammatory molecules under the regulation of NF-κB pathway such asprostaglandins produced from COX-2, ICAM-1 and MMP-1 and 3 etc. Also,the compounds have ability to inhibit cell proliferation under theregulation of AP-1 pathway through the inhibition of MEK1. In addition,the compounds have ability to inhibit angiogenesis mainly based on theinhibitory activities on VEGFr and PDGFr kinases. Thus, the compounds ofthe invention, and pharmaceutical compositions thereof, are useful asanti-inflammatory and/or immunosuppressive agents for the treatment ofvarious inflammatory diseases, and abnormal cell proliferation or asantiangiogenesis agents for the treatment of cancer. In certainembodiments, the compounds of the present invention can be used for thetreatment of diseases and disorders including, but not limited tosepsis, rheumatoid arthritis, psoriatic arthritis, osteoarthritis,inflammatory bowel disease (Crohn's disease and ulcerative colitis),multiple sclerosis, atopic dermatitis, psoriasis, asthma, osteoporosis,allergic rhinitis, ocular inflammation, hepatitis, autoimmune disorders,systemic lupus erthematosus, allograft rejection/graft versus hostdisease, diabetes, AIDS, solid tumor cancers, leukemia, lymphomas,non-hodgkin's B-cell lymphomas, chronical lymphocytic leukemia (CLL),multiple myeloma, eczema, urticaria, myasthenia gravis, idiopathicthrombocytopenia purpura, cardiovascular disease (e.g., myocardialinfarction, atherosclerosis), hepatitis, glomerulonephropathy,productive nephritis, adenovirus, diseases/disorders of the centralnervous system (e.g., stroke, Alzheimer's disease, epilepsy) and for thetreatment of the symptoms of malaria, to name a few. The inventivecompounds also find use in the prevention of restenosis of blood vesselssubject to traumas such as angioplasty and stenting.

In addition, it has been shown that photoaging of undamaged skin due toUVB irradiation exposure is inhibited by administering an agent thatinhibits one or both of the transcription factors AP-1 and NF-κB to theskin prior to such exposure (See, for example, U.S. Pat. No. 5,837,224).Therefore, the inventive compounds, and pharmaceutical compositionsthereof, are useful in the treatment of photoaging-relateddisorders/conditions.

1) General Description of Compounds of the Invention

In certain embodiments, the compounds of the invention include compoundsof the general formula (I) as further defined below:

wherein R₁ is hydrogen, aliphatic, heteroaliphatic, alicyclic,heteroalicyclic, aryl or heteroaryl;

R₂ and R₃ are each independently hydrogen, halogen, hydroxyl, protectedhydroxyl, or an aliphatic, heteroaliphatic, alicyclic, heteroalicyclic,aryl or heteroaryl moiety; or

R₁ and R₂, when taken together, may form a substituted or unsubstituted,saturated or unsaturated cyclic ring of 3 to 8 carbon atoms; or

R₁ and R₃, when taken together, may form a substituted or unsubstituted,saturated or unsaturated cyclic ring of 3 to 8 carbon atoms;

R₄ is hydrogen or halogen;

R₅ is hydrogen, an oxygen protecting group or a prodrug;

R₆ is hydrogen, hydroxyl, or protected hydroxyl;

n is 0-2;

R₇, for each occurrence, is independently hydrogen, hydroxyl, orprotected hydroxyl;

R₁ is hydrogen, halogen, hydroxyl, protected hydroxyl, alkyloxy, or analiphatic moiety optionally substituted with hydroxyl, protectedhydroxyl, SR₁₂, or NR₁₂R₁₃;

R₉ is hydrogen, halogen, hydroxyl, protected hydroxyl, OR₁₂, SR₁₂,NR₁₂R₁₃, —X₁(CH₂)_(p)X₂—R₁₄, or is lower alkyl optionally substitutedwith hydroxyl, protected hydroxyl, halogen, amino, protected amino, or—X₁(CH₂)_(p)X₂—R₁₄;

-   -   wherein R₁₂ and R₁₃ are, independently for each occurrence,        hydrogen, aliphatic, heteroaliphatic, alicyclic,        heteroalicyclic, aryl or heteroaryl; or a protecting group, or        R₁₂ and R₁₃, taken together may form a saturated or unsaturated        cyclic ring containing 1 to 4 carbon atoms and 1 to 3 nitrogen        or oxygen atoms, and each of R₁₂ and R₁₃ are optionally further        substituted with one or more occurrences of hydroxyl, protected        hydroxyl, alkyloxy, amino, protected amino, alkylamino,        aminoalkyl, or halogen,    -   wherein X₁ and X₂ are each independently absent, or are oxygen,        NH, or —N(alkyl), or wherein X₂—R₁₄ together are N₃ or are a        saturated or unsaturated heterocyclic moiety,    -   p is 2-10, and    -   R₁₄ is hydrogen, or an aryl, heteroaryl, alkylaryl, or        alkylheteroaryl moiety, or is —(C═O)NHR₁₅—(C═O)OR₁₅, or        —(C═O)R₁₅, wherein each occurrence of R₁₅ is independently        hydrogen, aliphatic, heteroaliphatic, alicyclic,        heteroalicyclic, aryl or heteroaryl; or R₁₄ is —SO₂(R₁₆),        wherein R₁₆ is an aliphatic moiety, wherein one or more of R₁₄,        R₁₅, or R₁₆ are optionally substituted with one or more        occurrences of hydroxyl, protected hydroxyl, alkyloxy, amino,        protected amino, alkylamino, aminoalkyl, or halogen; or

R₈ and R₉ may, when taken together, form a saturated or unsaturatedcyclic ring containing 1 to 4 carbon atoms and 1 to 3 nitrogen or oxygenatoms and is optionally substituted with hydroxyl, protected hydroxyl,alkyloxy, amino, protected amino, alkylamino, aminoalkyl, or halogen;

R₁₀ is hydrogen, hydroxyl, protected hydroxyl, amino, or protectedamino;

R₁₁ is hydrogen, hydroxyl or protected hydroxyl;

X is absent or is O, NH, N-alkyl, CH₂ or S;

Y is CHR₁₇, O, C═O, CR₁₇ or NR₁₇; and Z is CHR₁₈, O, C═O, CR₁₈ or NR₁₈,wherein each occurrence of R₁₇ and R₁₈ is independently hydrogen oraliphatic, or R₁₇ and R₁₈ taken together is —O—, —CH₂— or —NR₁₉—,wherein R₁₉ is hydrogen or lower alkyl, and Y and Z may be connected bya single or double bond; and

pharmaceutically acceptable derivatives thereof.

In certain embodiments of compounds described directly above andcompounds as described in certain classes and subclasses herein, thefollowing groups do not occur simultaneously as defined:

X is oxygen,

R₁ is methyl with S-configuration,

R₂ and R₃ are each hydrogen,

R₄ is hydrogen,

R₅ is hydrogen, lower alkyl or lower alkanoyl,

R₆ is OR′, where R′ is hydrogen, lower alkyl or lower alkanoyl withS-configuration,

R₇ is hydrogen,

Y and Z together represent —CHR₁₇—CHR₁₈— or —CR₁₇═CR₁₈—, wherein

R₁₇ and R₁₈ are independently hydrogen, or when Y and Z are—CHR₁₇—CHR₁₈, R₁₇ and R₁₈ taken together are —O—;

R₈ is hydrogen or OR′, where R′ is hydrogen, lower alkyl or loweralkanoyl,

R₉ is OR′, where R′ is hydrogen, lower alkyl or lower alkanoyl,

R₁₀ is OR″, where R″ is hydrogen, lower alkyl or lower alkanoyl; and

R¹¹ is hydrogen.

In certain other embodiments, compounds of formula (I) are defined asfollows:

R₁ is hydrogen, straight or branched lower alkyl, straight or branchedlower heteroalkyl, or aryl,

-   -   wherein the alkyl, heteroalkyl, and aryl groups may optionally        be substituted with one or more occurrences of halogen, hydroxyl        or protected hydroxyl;

R₂ and R₃ are each independently hydrogen, halogen, hydroxyl, protectedhydroxyl, straight or branched lower alkyl, straight or branched lowerheteroalkyl, or aryl,

-   -   wherein the alkyl, heteroalkyl, and aryl groups may optionally        be substituted with one or more occurrences of halogen, hydroxyl        or protected hydroxyl; or

R₁ and R₂, when taken together, may form a saturated or unsaturatedcyclic ring of 3 to 8 carbon atoms, optionally substituted with one ormore occurrences of halogen; or

R₁ and R₃, when taken together, may form a saturated or unsaturatedcyclic ring of 3 to 8 carbon atoms, optionally substituted with one ormore occurrences of halogen;

R₄ is hydrogen or halogen;

R₅ is hydrogen or a protecting group;

R₆ is hydrogen, hydroxyl, or protected hydroxyl;

n is 0-2;

R₇, for each occurrence, is independently hydrogen, hydroxyl, orprotected hydroxyl;

R₈ is hydrogen, halogen, hydroxyl, protected hydroxyl, alkyloxy, orlower alkyl optionally substituted with hydroxyl, protected hydroxyl,SR₁₂, or NR₁₂R₁₃;

R₉ is hydrogen, halogen, hydroxyl, protected hydroxyl, OR₁₂, SR₁₂,NR₁₂R₁₃, —X₁(CH₂)_(p)X₂—R₁₄, or is lower alkyl optionally substitutedwith hydroxyl, protected hydroxyl, halogen, amino, protected amino, or—X₁(CH₂)_(p)X₂—R₁₄;

-   -   wherein R₁₂ and R₁₃ are, independently for each occurrence,        hydrogen, lower alkyl, aryl, heteroaryl, alkylaryl, or        alkylheteroaryl, or a protecting group, or R₁₂ and R₁₃, taken        together may form a saturated or unsaturated cyclic ring        containing 1 to 4 carbon atoms and 1 to 3 nitrogen or oxygen        atoms, and each of R₁₂ and R₁₃ are optionally further        substituted with one or more occurrences of hydroxyl, protected        hydroxyl, alkyloxy, amino, protected amino, alkylamino,        aminoalkyl, or halogen,    -   wherein X₁ and X₂ are each independently absent, or are oxygen,        NH, or —N(alkyl), or wherein X₂—R₁₄ together are N₃ or are a        saturated or unsaturated heterocyclic moiety,    -   p is 2-10, and

R₁₄ is hydrogen, or an aryl, heteroaryl, alkylaryl, or alkylheteroarylmoiety, or is —(C═O)NHR₁₅—(C═O)OR₁₅, or —(C═O)R₁₅, wherein eachoccurrence of R₁₅ is independently hydrogen, alkyl, heteroalkyl, aryl,heteroaryl, alkylaryl, or alkylheteroaryl, or R₁₄ is —SO₂(R₁₆), whereinR₁₆ is an alkyl moiety, wherein one or more of R₁₄, R₁₅, or R₁₆ areoptionally substituted with one or more occurrences of hydroxyl,protected hydroxyl, alkyloxy, amino, protected amino, alkylamino,aminoalkyl, or halogen; or

R₈ and R₉ may, when taken together, form a saturated or unsaturatedcyclic ring containing 1 to 4 carbon atoms and 1 to 3 nitrogen or oxygenatoms and is optionally substituted with hydroxyl, protected hydroxyl,alkyloxy, amino, protected amino, alkylamino, aminoalkyl, or halogen;

R₁₀ is hydrogen, hydroxyl, protected hydroxyl, amino, or protectedamino;

R₁₁ is hydrogen, hydroxyl or protected hydroxyl;

X is absent or is O, NH, N-alkyl, CH₂ or S;

Y is CHR₁₇, O, C═O, CR₁₇ or NR₁₇; and Z is CHR₁₈, O, C═O, CR₁₈ or NR₁₈,wherein each occurrence of R₁₇ and R₁₈ is independently hydrogen orlower alkyl, or R₁₇ and R₁₈ taken together is —O—, —CH₂— or —NR₁₉—,wherein R₁₉ is hydrogen or lower alkyl, and Y and Z may be connected bya single or double bond; and

pharmaceutically acceptable derivatives thereof.

In certain other embodiments, compounds of formula (I) are defined asfollows:

R₁ is hydrogen, straight or branched lower alkyl, straight or branchedlower heteroalkyl, or aryl,

-   -   wherein the alkyl, heteroalkyl, and aryl groups may optionally        be substituted with one or more occurrences of halogen, hydroxyl        or protected hydroxyl;

R₂ and R₃ are each independently hydrogen, halogen, hydroxyl, protectedhydroxyl, straight or branched lower alkyl, straight or branched lowerheteroalkyl, or aryl,

-   -   wherein the alkyl, heteroalkyl, and aryl groups may optionally        be substituted with one or more occurrences of halogen, hydroxyl        or protected hydroxyl; or

R₁ and R₂, when taken together, may form a saturated or unsaturatedcyclic ring of 3 to 8 carbon atoms, optionally substituted with one ormore occurrences of halogen;

R₄ is hydrogen or halogen;

R₅ is hydrogen or a protecting group;

R₆ is hydrogen, hydroxyl, or protected hydroxyl;

n is 0-2;

R₇, for each occurrence, is independently hydrogen, hydroxyl, orprotected hydroxyl;

R₈ is hydrogen, halogen, hydroxyl, protected hydroxyl, alkyloxy, orlower alkyl optionally substituted with hydroxyl or protected hydroxyl;

R₉ is hydrogen, halogen, hydroxyl, protected hydroxyl, OR₁₂, NR₁₂R₁₃,—X₁(CH₂)_(p)X₂—R₁₄, or is lower alkyl optionally substituted withhydroxyl, protected hydroxyl, halogen, amino, protected amino, or—X₁(CH₂)_(p)X₂—R₁₄;

-   -   wherein R₁₂ and R₁₃ are, independently for each occurrence,        hydrogen, lower alkyl, aryl, heteroaryl, alkylaryl, or        alkylheteroaryl, or a protecting group, or R₁₂ and R₁₃, taken        together may form a saturated or unsaturated cyclic ring        containing 1 to 4 carbon atoms and 1 to 3 nitrogen or oxygen        atoms, and each of R₁₂ and R₁₃ are optionally further        substituted with one or more occurrences of hydroxyl, protected        hydroxyl, alkyloxy, amino, protected amino, alkylamino,        aminoalkyl, or halogen,    -   wherein X₁ and X₂ are each independently absent, or are oxygen,        NH, or —N(alkyl), or wherein X₂—R₁₄ together are N₃ or are a        saturated or unsaturated heterocyclic moiety,    -   p is 2-10, and    -   R₁₄ is hydrogen, or an aryl, heteroaryl, alkylaryl, or        alkylheteroaryl moiety, or is —(C═O)NHR₁₅—(C═O)OR₁₅, or        —(C═O)R₁₅, wherein each occurrence of R₁₅ is independently        hydrogen, alkyl, heteroalkyl, aryl, heteroaryl, alkylaryl, or        alkylheteroaryl, or R₁₄ is —SO₂(R₁₆), wherein R₁₆ is an alkyl        moiety, wherein one or more of R₁₄, R₁₅, or R₁₆ are optionally        substituted with one or more occurrences of hydroxyl, protected        hydroxyl, alkyloxy, amino, protected amino, alkylamino,        aminoalkyl, or halogen; or

R₈ and R₉ may, when taken together, form a saturated or unsaturatedcyclic ring containing 1 to 4 carbon atoms and 1 to 3 nitrogen or oxygenatoms and is optionally substituted with hydroxyl, protected hydroxyl,alkyloxy, amino, protected amino, alkylamino, aminoalkyl, or halogen;

R₁₀ is hydrogen, hydroxyl, protected hydroxyl, amino, or a protectedamino group;

R₁₁ is hydrogen, hydroxyl, or protected hydroxyl;

X is absent or is O, NH, or CH₂;

Y is —CHR₁₇, O, C═O, CR₁₇ or NR₁₇, and Z is CHR₁₅, O, C═O, CR₁₈ or NR₁₈,wherein each occurrence of R₁₇ and R₁₈ is independently hydrogen orlower alkyl; and

pharmaceutically acceptable derivatives thereof.

In certain embodiments, the present invention defines certain classes ofcompounds which are of special interest. For example, one class ofcompounds of special interest includes those compounds having thestructure of formula (I) in which X is O, and n is 1 and the compoundhas the structure:

wherein R₁-R₁₁, Y and Z are as previously defined.

Another class of compounds of special interest includes compounds havingthe structure of formula (I) in which R₄ is halogen (Hal), and thecompound has the structure:

wherein R₁-R₃, R₅-R₁₁, X, Y, Z and n are as previously defined, andwherein Hal is a halogen selected from fluorine, bromine, chlorine andiodine.

Another class of compounds of special interest includes compounds havingthe structure of formula (I) in which Y and Z together represent—CH═CH—, and the compound has the structure:

wherein R₁-R₁₁, X and n are as previously defined.

Another class of compounds of special interest includes compounds havingthe structure of formula (I) in which R₁ and R₂ are each methyl, and R₃is hydrogen and the compound has the structure:

wherein R₄-R₁₁, n, X, Y and Z are as previously defined.

Another class of compounds of special interest includes compounds havingthe structure of formula (I) in which R₉ is NR₁₂R₁₃, and the compoundhas the structure:

wherein R₁-R₁₃, n, X, Y and Z are as previously defined,

and R₁₃ and R₈ may additionally, when taken together, form a saturatedor unsaturated cyclic ring containing 1 to 4 carbon atoms and 1 to 3nitrogen or oxygen atoms and is optionally substituted with hydroxyl,protected hydroxyl, alkyloxy, amino, protected amino, alkylamino,aminoalkyl, and halogen.

Another class of compounds of special interest includes compounds havingthe structure of formula (I) in which R₉ is OR₁₂, and the compound hasthe structure:

wherein R₁-R₁₂, n, X, Y and Z are as previously defined.

Another class of compounds of special interest includes compounds havingthe structure of formula (I) in which R₉ is —X₁(CH₂)_(p)X₂R₁₄, and thecompound has the structure:

wherein R₁-R₁₁, R₁₄, X, Y, Z, n, X₁, X₂ and p are as defined above.

The following structures illustrate several exemplary types of compoundsof these classes. Additional compounds are described in theExemplification herein. Other compounds of the invention will be readilyapparent to the reader:

A number of important subclasses of each of the foregoing classesdeserve separate mention; these subclasses include subclasses of theforegoing classes in which:

i) R₁ is hydrogen, aryl or lower alkyl;

ii) R₁ is hydrogen, phenyl, methyl or ethyl;

iii) R₁ is methyl;

iv) R₂ is hydrogen, halogen or lower alkyl;

v) R₂ is hydrogen, F, methyl or ethyl;

vi) R₂ is methyl;

vii) R₃ is hydrogen;

viii) R₁ and R₂ are each methyl and R₃ is hydrogen;

ix) R₁ and R₂, taken together, form a 5- to 6-membered cycloalkylmoiety;

x) R₁ and R₃, taken together, form a 5- to 6-membered cycloalkyl moiety;

xi) R₄ is a halogen selected from fluorine, chlorine, bromine, andiodine;

xii) R₄ is a hydrogen;

xiii) R₄ is fluorine;

xiv) R₅ is a protecting group, hydrogen or a prodrug moiety;

xv) R₅ is an oxygen protecting group;

xvi) R₅ is an oxygen protecting group selected from methyl ether,substituted methyl ether, substituted ethyl ether, substituted benzylether, silyl ether, ester, carbonate, cyclic acetal and ketal;

xvii) R₆ is hydrogen, hydroxyl or protected hydroxyl;

xviii) R₆ is protected hydroxyl and the protecting group is an oxygenprotecting group;

xix) R₆ is protected hydroxyl and the protecting group is an oxygenprotecting group selected from methyl ether, substituted methyl ether,substituted ethyl ether, substituted benzyl ether, silyl ether, ester,carbonate, cyclic acetal and ketal;

xx) R₆ is protected hydroxyl and the protecting group is a prodrugmoiety;

xxi) n is 1;

xxii) R₇ is hydrogen;

xxiii) R₇ is hydroxyl;

xxiv) R₇ is protected hydroxyl and the protecting group is an oxygenprotecting group;

xxv) R₇ is a protected hydroxyl and the protecting group is an oxygenprotecting group selected from methyl ether, substituted methyl ether,substituted ethyl ether, substituted benzyl ether, silyl ether, ester,carbonate, cyclic acetal and ketal;

xxvi) R₇ is protected hydroxyl and the protecting group is a prodrugmoiety;

xxvii) Y and Z together represent —CH═CH—;

xxviii) Y and Z together represent trans —CH═CH—;

xxix) Y and Z together represent —CR₁₇═CR₁₈—;

xxx) Y and Z together represent trans CR₁₇═CR₁₈—;

xxxi) Y and Z together are an epoxide;

xxxii) Y and Z together are an aziridine;

xxxiii) Y and Z together are cyclopropyl;

xxxiv) Y and Z together are —CH₂—CH₂—;

xxxv) Z is O;

xxxvi) Y is O;

xxxvii) Z is C═O and Y is CHR₁₇;

xxxviii) Z is NR₁₈ and Y is CHR₁₇;

xxxix) Z is CHR₁₈ and Y is C═O;

xl) Z is CHR₁₈ and Y is NR₁₇;

xli) X is O or NH;

xlii) R₈ is hydrogen;

xliii) R₈ is halogen, hydroxyl, protected hydroxyl, alkyloxy, or loweralkyl optionally substituted when one or more hydroxyl or protectedhydroxyl groups;

xliv) R₉ is hydrogen;

xlv) R₉ is OR₁₂, wherein R₁₂ is methyl, ethyl, propyl, isopropyl, butyl,—CH₂COOMe, Bn, PMB (MPM), 3,4-CIBn, or R₉ is

xlvi) R₉ is NR₁₂R₁₃, wherein R₁₂ is methyl, ethyl, propyl, isopropyl, orbutyl, optionally substituted with one or more occurrences of hydroxylor protected hydroxyl, and R₁₃ is hydrogen or lower alkyl, or NR₁₂R¹³together represents a 5- or 6-membered heterocyclic moiety;

xlvii) R₉ is O(CH₂)_(p)X₂R₁₄, wherein X₂R₁₄ together represent N₃, NMe₂,NHAc, NHSO₂Me, NHCONHMe, NHCONHPh, morpholine, imidazole, aminopyridine,or any one of:

xlviii) R₁₀ is hydroxyl or protected hydroxyl;

xlix) R₁₀ is hydroxyl; and/or

l) R₁₁ is hydrogen.

As the reader will appreciate, compounds of particular interest include,among others, those which share the attributes of one or more of theforegoing subclasses. Some of those subclasses are illustrated by thefollowing sorts of compounds:

I) Compounds of the Formula (and Pharmaceutically Acceptable DerivativesThereof):

wherein R₅-R₈, R₁₀-R₁₃ are as defined above and in subclasses herein.

II) Compounds of the Formula (and Pharmaceutically AcceptableDerivatives Thereof):

wherein R₅-R₈, R₁₀-R₁₃ are as defined above and in subclasses herein.

III) Compounds of the Formula (and Pharmaceutically AcceptableDerivatives Thereof):

wherein R₅-R₈, R₁₀, and R₁₂ are as defined above and in subclassesherein.

IV) Compounds of the Formula (and Pharmaceutically AcceptableDerivatives Thereof):

wherein R₅-R₈, R₁₀ and R₁₂ are as defined above and in subclassesherein.

V) Compounds of the Formula (and Pharmaceutically Acceptable DerivativesThereof):

wherein R₅-R₈, R₁₀, R₁₄, X₁, X₂ and p are as defined above and insubclasses herein.

VI) Compounds of the Formula (and Pharmaceutically AcceptableDerivatives Thereof):

wherein R₅-R₈, R₁₀, R₁₄, X₁, X₂ and p are as defined above and insubclasses herein.

It will also be appreciated that for each of the subgroups I-VIdescribed above, a variety of other subclasses are of special interest,including, but not limited to those classes described above 0-1) andclasses, subclasses and species of compounds described above and in theexamples herein.

Some of the foregoing compounds can comprise one or more asymmetriccenters, and thus can exist in various isomeric forms, e.g.,stereoisomers and/or diastereomers. Thus, inventive compounds andpharmaceutical compositions thereof may be in the form of an individualenantiomer, diastereomer or geometric isomer, or may be in the form of amixture of stereoisomers. In certain embodiments, the compounds of theinvention are enantiopure compounds. In certain other embodiments,mixtures of stereoisomers or diastereomers are provided.

Furthermore, certain compounds, as described herein may have one or moredouble bonds that can exist as either the Z or E isomer, unlessotherwise indicated. The invention additionally encompasses thecompounds as individual isomers substantially free of other isomers andalternatively, as mixtures of various isomers, e.g., racemic mixtures ofstereoisomers. In addition to the above-mentioned compounds per se, thisinvention also encompasses pharmaceutically acceptable derivatives ofthese compounds and compositions comprising one or more compounds of theinvention and one or more pharmaceutically acceptable excipients oradditives.

Compounds of the invention may be prepared by crystallization ofcompound of formula (I) under different conditions and may exist as oneor a combination of polymorphs of compound of general formula (I)forming part of this invention. For example, different polymorphs may beidentified and/or prepared using different solvents, or differentmixtures of solvents for recrystallization; by performingcrystallizations at different temperatures; or by using various modes ofcooling, ranging from very fast to very slow cooling duringcrystallizations. Polymorphs may also be obtained by heating or meltingthe compound followed by gradual or fast cooling. The presence ofpolymorphs may be determined by solid probe NMR spectroscopy, IRspectroscopy, differential scanning calorimetry, powder X-raydiffractogram and/or other techniques. Thus, the present inventionencompasses inventive compounds, their derivatives, their tautomericforms, their stereoisomers, their polymorphs, their pharmaceuticallyacceptable salts their pharmaceutically acceptable solvates andpharmaceutically acceptable compositions containing them.

2) Compounds and Definitions

As discussed above, this invention provides novel compounds with a rangeof biological properties. Compounds of this invention have biologicalactivities relevant for the treatment of inflammatory and immunedisorders, photoaging and cancer. In certain embodiments, the compoundsof the invention are useful for the treatment of rheumatoid arthritis,psoriasis, Multiple sclerosis, and asthma. In certain other embodiments,the inventive compounds also find use in the prevention of restenosis ofblood vessels subject to traumas such as angioplasty and stenting.

Compounds of this invention include those specifically set forth aboveand described herein, and are illustrated in part by the variousclasses, subgenera and species disclosed elsewhere herein.

Additionally, the present invention provides pharmaceutically acceptablederivatives of the inventive compounds, and methods of treating asubject using these compounds, pharmaceutical compositions thereof, oreither of these in combination with one or more additional therapeuticagents. The phrase, “pharmaceutically acceptable derivative”, as usedherein, denotes any pharmaceutically acceptable salt, ester, or salt ofsuch ester, of such compound, or any other adduct or derivative which,upon administration to a patient, is capable of providing (directly orindirectly) a compound as otherwise described herein, or a metabolite orresidue thereof. Pharmaceutically acceptable derivatives thus includeamong others pro-drugs. A pro-drug is a derivative of a compound,usually with significantly reduced pharmacological activity, whichcontains an additional moiety, which is susceptible to removal in vivoyielding the parent molecule as the pharmacologically active species. Anexample of a pro-drug is an ester, which is cleaved in vivo to yield acompound of interest. Pro-drugs of a variety of compounds, and materialsand methods for derivatizing the parent compounds to create thepro-drugs, are known and may be adapted to the present invention.Certain exemplary pharmaceutical compositions and pharmaceuticallyacceptable derivatives will be discussed in more detail herein below.

Certain compounds of the present invention, and definitions of specificfunctional groups are also described in more detail below. For purposesof this invention, the chemical elements are identified in accordancewith the Periodic Table of the Elements, CAS version, Handbook ofChemistry and Physics, 75^(th) Ed., inside cover, and specificfunctional groups are generally defined as described therein.Additionally, general principles of organic chemistry, as well asspecific functional moieties and reactivity, are described in “OrganicChemistry”, Thomas Sorrell, University Science Books, Sausalito: 1999,the entire contents of which are incorporated herein by reference.Furthermore, it will be appreciated by one of ordinary skill in the artthat the synthetic methods, as described herein, utilize a variety ofprotecting groups. By the term “protecting group”, has used herein, itis meant that a particular functional moiety, e.g., O, S, or N, istemporarily blocked so that a reaction can be carried out selectively atanother reactive site in a multifunctional compound. In preferredembodiments, a protecting group reacts selectively in good yield to givea protected substrate that is stable to the projected reactions; theprotecting group must be selectively removed in good yield by readilyavailable, preferably nontoxic reagents that do not attack the otherfunctional groups; the protecting group forms an easily separablederivative (more preferably without the generation of new stereogeniccenters); and the protecting group has a minimum of additionalfunctionality to avoid further sites of reaction. As detailed herein,oxygen, sulfur, nitrogen and carbon protecting groups may be utilized.For example, in certain embodiments, as detailed herein, certainexemplary oxygen protecting groups are utilized. These oxygen protectinggroups include, but are not limited to methyl ethers, substituted methylethers (e.g., MOM (methoxymethyl ether), MTM (methylthiomethyl ether),BOM (benzyloxymethyl ether), PMBM or MPM (p-methoxybenzyloxymethylether), to name a few), substituted ethyl ethers, substituted benzylethers, silyl ethers (e.g., TMS (trimethylsilyl ether), TES(triethylsilylether), TIPS (triisopropylsilyl ether), TBDMS(t-butyldimethylsilyl ether), tribenzyl silyl ether, TBDPS(t-butyldiphenyl silyl ether), to name a few), esters (e.g., formate,acetate, benzoate (Bz), trifluoroacetate, dichloroacetate, to name afew), carbonates, cyclic acetals and ketals. In certain other exemplaryembodiments, nitrogen protecting groups are utilized. These nitrogenprotecting groups include, but are not limited to, carbamates (includingmethyl, ethyl and substituted ethyl carbamates (e.g., Troc), to name afew) amides, cyclic imide derivatives, N-Alkyl and N-Aryl amines, iminederivatives, and enamine derivatives, to name a few. Certain otherexemplary protecting groups are detailed herein, however, it will beappreciated that the present invention is not intended to be limited tothese protecting groups; rather, a variety of additional equivalentprotecting groups can be readily identified using the above criteria andutilized in the present invention. Additionally, a variety of protectinggroups are described in “Protective Groups in Organic Synthesis” ThirdEd. Greene, T. W. and Wuts, P. G., Eds., John Wiley & Sons, New York:1999, the entire contents of which are hereby incorporated by reference.

It will be appreciated that the compounds, as described herein, may besubstituted with any number of substituents or functional moieties. Ingeneral, the term “substituted” whether preceded by the term“optionally” or not, and substituents contained in formulas of thisinvention, refer to the replacement of hydrogen radicals in a givenstructure with the radical of a specified substituent. When more thanone position in any given structure may be substituted with more thanone substituent selected from a specified group, the substituent may beeither the same or different at every position. As used herein, the term“substituted” is contemplated to include all permissible substituents oforganic compounds. In a broad aspect, the permissible substituentsinclude acyclic and cyclic, branched and unbranched, carbocyclic andheterocyclic, aromatic and nonaromatic substituents of organiccompounds. For purposes of this invention, heteroatoms such as nitrogenmay have hydrogen substituents and/or any permissible substituents oforganic compounds described herein which satisfy the valencies of theheteroatoms. Furthermore, this invention is not intended to be limitedin any manner by the permissible substituents of organic compounds.Combinations of substituents and variables envisioned by this inventionare preferably those that result in the formation of stable compoundsuseful in the treatment, for example of inflammatory and proliferativedisorders, including, but not limited to rheumatoid arthritis,psoriasis, asthma and cancer. The term “stable”, as used herein,preferably refers to compounds which possess stability sufficient toallow manufacture and which maintain the integrity of the compound for asufficient period of time to be detected and preferably for a sufficientperiod of time to be useful for the purposes detailed herein.

The term “aliphatic”, as used herein, includes both saturated andunsaturated, straight chain (i.e., unbranched) or branched aliphatichydrocarbons, which are optionally substituted with one or morefunctional groups. As will be appreciated by one of ordinary skill inthe art, “aliphatic” is intended herein to include, but is not limitedto, alkyl, alkenyl, alkynyl moieties. Thus, as used herein, the term“alkyl” includes straight and branched alkyl groups. An analogousconvention applies to other generic terms such as “alkenyl”, “alkynyl”and the like. Furthermore, as used herein, the terms “alkyl”, “alkenyl”,“alkynyl” and the like encompass both substituted and unsubstitutedgroups. In certain embodiments, as used herein, “lower alkyl” is used toindicate those alkyl groups (cyclic, acyclic, substituted,unsubstituted, branched or unbranched) having 1-6 carbon atoms.

In certain embodiments, the alkyl, alkenyl and alkynyl groups employedin the invention contain 1-20 aliphatic carbon atoms. In certain otherembodiments, the alkyl, alkenyl, and alkynyl groups employed in theinvention contain 1-10 aliphatic carbon atoms. In yet other embodiments,the alkyl, alkenyl, and alkynyl groups employed in the invention contain1-8 aliphatic carbon atoms. In still other embodiments, the alkyl,alkenyl, and alkynyl groups employed in the invention contain 1-6aliphatic carbon atoms. In yet other embodiments, the alkyl, alkenyl,and alkynyl groups employed in the invention contain 1-4 carbon atoms.Illustrative aliphatic groups thus include, but are not limited to, forexample, methyl, ethyl, n-propyl, isopropyl, allyl, n-butyl, sec-butyl,isobutyl, tert-butyl, n-pentyl, sec-pentyl, isopentyl, tert-pentyl,n-hexyl, sec-hexyl, moieties and the like, which again, may bear one ormore substituents. Alkenyl groups include, but are not limited to, forexample, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, and thelike. Representative alkynyl groups include, but are not limited to,ethynyl, 2-propynyl(propargyl), 1-propynyl and the like.

The term “alicyclic”, as used herein, refers to compounds which combinethe properties of aliphatic and cyclic compounds and include but are notlimited to cyclic, or polycyclic aliphatic hydrocarbons and bridgedcycloalkyl compounds, which are optionally substituted with one or morefunctional groups. As will be appreciated by one of ordinary skill inthe art, “alicyclic” is intended herein to include, but is not limitedto, cycloalkyl, cycloalkenyl, and cycloalkynyl moieties, which areoptionally substituted with one or more functional groups. Illustrativealicyclic groups thus include, but are not limited to, for example,cyclopropyl, —CH₂-cyclopropyl, cyclobutyl, —CH₂-cyclobutyl, cyclopentyl,—CH₂-cyclopentyl-n, cyclohexyl, —CH₂-cyclohexyl, cyclohexenylethyl,cyclohexanylethyl, norborbyl moieties and the like, which again, maybear one or more substituents.

The term “alkoxy” (or “alkyloxy”), or “thioalkyl” as used herein refersto an alkyl group, as previously defined, attached to the parentmolecular moiety through an oxygen atom or through a sulfur atom. Incertain embodiments, the alkyl group contains 1-20 aliphatic carbonatoms. In certain other embodiments, the alkyl group contains 1-10aliphatic carbon atoms. In yet other embodiments, the alkyl, alkenyl,and alkynyl groups employed in the invention contain 1-8 aliphaticcarbon atoms. In still other embodiments, the alkyl group contains 1-6aliphatic carbon atoms. In yet other embodiments, the alkyl groupcontains 1-4 aliphatic carbon atoms. Examples of alkoxy, include but arenot limited to, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy,tert-butoxy, neopentoxy and n-hexoxy. Examples of thioalkyl include, butare not limited to, methylthio, ethylthio, propylthio, isopropylthio,n-butylthio, and the like.

The term “alkylamino” refers to a group having the structure —NHR′wherein R′ is alkyl, as defined herein. The term “aminoalkyl” refers toa group having the structure NH₂R′—, wherein R′ is alkyl, as definedherein. In certain embodiments, the alkyl group contains 1-20 aliphaticcarbon atoms. In certain other embodiments, the alkyl group contains1-10 aliphatic carbon atoms. In yet other embodiments, the alkyl,alkenyl, and alkynyl groups employed in the invention contain 1-8aliphatic carbon atoms. In still other embodiments, the alkyl groupcontains 1-6 aliphatic carbon atoms. In yet other embodiments, the alkylgroup contains 1-4 aliphatic carbon atoms. Examples of alkylaminoinclude, but are not limited to, methylamino, ethylamino,iso-propylamino and the like.

Some examples of substituents of the above-described aliphatic (andother) moieties of compounds of the invention include, but are notlimited to aliphatic; heteroaliphatic; aryl; heteroaryl; alkylaryl;alkylheteroaryl; alkoxy; aryloxy; heteroalkoxy; heteroaryloxy;alkylthio; arylthio; heteroalkylthio; heteroarylthio; F; Cl; Br; I; —OH;—NO₂; —CN; —CF₃; —CH₂CF₃; —CHCl₂; —CH₂OH; —CH₂CH₂OH; —CH₂NH₂;—CH₂SO₂CH₃; —C(O)R_(x); —CO₂(R_(x)); —CON(R_(x))₂; —OC(O)R_(x);—OCO₂R_(x); —OCON(R_(x))₂; —N(R_(x))₂; —S(O)₂R_(x); —NR_(x)(CO)R_(x)wherein each occurrence of R_(x) independently includes, but is notlimited to, aliphatic, heteroaliphatic, aryl, heteroaryl, alkylaryl, oralkylheteroaryl, wherein any of the aliphatic, heteroaliphatic,alkylaryl, or alkylheteroaryl substituents described above and hereinmay be substituted or unsubstituted, branched or unbranched, cyclic oracyclic, and wherein any of the aryl or heteroaryl substituentsdescribed above and herein may be substituted or unsubstituted.Additional examples of generally applicable substituents are illustratedby the specific embodiments shown in the Examples that are describedherein.

In general, the terms “aryl” and “heteroaryl”, as used herein, refer tostable mono- or polycyclic, heterocyclic, polycyclic, andpolyheterocyclic unsaturated moieties having preferably 3-14 carbonatoms, each of which may be substituted or unsubstituted. It will alsobe appreciated that aryl and heteroaryl moieties, as defined herein maybe attached via an aliphatic, alicyclic, heteroaliphatic,heteroalicyclic, alkyl or heteroalkyl moiety and thus also include-(aliphatic)aryl, -(heteroaliphatic)aryl, -(aliphatic)heteroaryl,-(heteroaliphatic)heteroaryl, -(alkyl)aryl, -(heteroalkyl)aryl,-(heteroalkyl)aryl, and -(heteroalkyl)heteroaryl moieties. Thus, as usedherein, the phrases “aryl or heteroaryl” and “aryl, heteroaryl,-(aliphatic)aryl, -(heteroaliphatic)aryl, -(aliphatic)heteroaryl,-(heteroaliphatic)heteroaryl, -(alkyl)aryl, -(heteroalkyl)aryl,-(heteroalkyl)aryl, and -(heteroalkyl)heteroaryl” are interchangeable.Substituents include, but are not limited to, any of the previouslymentioned substitutents, i.e., the substituents recited for aliphaticmoieties, or for other moieties as disclosed herein, resulting in theformation of a stable compound. In certain embodiments of the presentinvention, “aryl” refers to a mono- or bicyclic carbocyclic ring systemhaving one or two aromatic rings including, but not limited to, phenyl,naphthyl, tetrahydronaphthyl, indanyl, indenyl and the like. In certainembodiments of the present invention, the term “heteroaryl”, as usedherein, refers to a cyclic aromatic radical having from five to ten ringatoms of which one ring atom is selected from S, O and N; zero, one ortwo ring atoms are additional heteroatoms independently selected from S,O and N; and the remaining ring atoms are carbon, the radical beingjoined to the rest of the molecule via any of the ring atoms, such as,for example, pyridyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl,imidazolyl, thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl, oxadiazolyl,thiophenyl, furanyl, quinolinyl, isoquinolinyl, and the like.

It will be appreciated that aryl and heteroaryl groups (includingbicyclic aryl groups) can be unsubstituted or substituted, whereinsubstitution includes replacement of one, two or three of the hydrogenatoms thereon independently with any one or more of the followingmoieties including, but not limited to: aliphatic; heteroaliphatic;aryl; heteroaryl; alkylaryl; alkylheteroaryl; alkoxy; aryloxy;heteroalkoxy; heteroaryloxy; alkylthio; arylthio; heteroalkylthio;heteroarylthio; F; Cl; Br; I; —OH; —NO₂; —CN; —CF₃; —CH₂CF₃; —CHCl₂;—CH₂OH; —CH₂CH₂OH; —CH₂NH₂; —CH₂SO₂CH₃; —C(O)R_(x); —CO₂(R_(x));—CON(R_(x))₂; —OC(O)R_(x); —OCO₂R_(x); —OCON(R_(x))₂; —N(R_(x))₂;—S(O)₂R_(x); —NR_(x)(CO)R_(x) wherein each occurrence of R_(x)independently includes, but is not limited to, aliphatic,heteroaliphatic, aryl, heteroaryl, alkylaryl, or alkylheteroaryl,wherein any of the aliphatic, heteroaliphatic, alkylaryl, oralkylheteroaryl substituents described above and herein may besubstituted or unsubstituted, branched or unbranched, cyclic or acyclic,and wherein any of the aryl or heteroaryl substituents described aboveand herein may be substituted or unsubstituted. Additional examples ofgenerally applicable substituents are illustrated by the specificembodiments shown in the Examples that are described herein.

The term “cycloalkyl”, as used herein, refers specifically to groupshaving three to seven, preferably three to ten carbon atoms. Suitablecycloalkyls include, but are not limited to cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl and the like, which, as in the caseof aliphatic, heteroaliphatic or heterocyclic moieties, may optionallybe substituted with substituents including, but not limited toaliphatic; heteroaliphatic; aryl; heteroaryl; alkylaryl;alkylheteroaryl; alkoxy; aryloxy; heteroalkoxy; heteroaryloxy;alkylthio; arylthio; heteroalkylthio; heteroarylthio; F; Cl; Br; I; —OH;—NO₂; —CN; —CF₃; —CH₂CF₃; —CHCl₂; —CH₂OH; —CH₂CH₂OH; —CH₂NH₂;—CH₂SO₂CH₃; —C(O)R_(x); —CO₂(R_(x)); —CON(R_(x))₂; —OC(O)R_(x);—OCO₂R_(x); —OCON(R_(x))₂; —N(R_(x))₂; —S(O)₂R_(x); —NR_(x)(CO)R_(x)wherein each occurrence of R_(x) independently includes, but is notlimited to, aliphatic, heteroaliphatic, aryl, heteroaryl, alkylaryl, oralkylheteroaryl, wherein any of the aliphatic, heteroaliphatic,alkylaryl, or alkylheteroaryl substituents described above and hereinmay be substituted or unsubstituted, branched or unbranched, cyclic oracyclic, and wherein any of the aryl or heteroaryl substituentsdescribed above and herein may be substituted or unsubstituted.Additional examples of generally applicable substituents are illustratedby the specific embodiments shown in the Examples that are describedherein.

The term “heteroaliphatic”, as used herein, refers to aliphatic moietiesin which one or more carbon atoms in the main chain have beensubstituted with a heteroatom. Thus, a heteroaliphatic group refers toan aliphatic chain which contains one or more oxygen, sulfur, nitrogen,phosphorus or silicon atoms, e.g., in place of carbon atoms.Heteroaliphatic moieties may be branched or linear unbranched. Incertain embodiments, heteroaliphatic moieties are substituted byindependent replacement of one or more of the hydrogen atoms thereonwith one or more moieties including, but not limited to aliphatic;alicyclic; heteroaliphatic; heteroalicyclic; aryl; heteroaryl;alkylaryl; alkylheteroaryl; alkoxy; aryloxy; heteroalkoxy;heteroaryloxy; alkylthio; arylthio; heteroalkylthio; heteroarylthio; F;Cl; Br; I; —OH; —NO₂; —CN; —CF₃; —CH₂CF₃; —CHCl₂; —CH₂OH; —CH₂CH₂OH;—CH₂NH₂; —CH₂SO₂CH₃; —C(O)R_(x); —CO₂(R_(x)); —CON(R_(x))₂; —OC(O)R_(x);—OCO₂R_(x); —OCON(R_(x))₂; —N(R_(x))₂; —S(O)₂R_(x); —NR—(CO)R_(x)wherein each occurrence of R_(x) independently includes, but is notlimited to, aliphatic, alicyclic, heteroaliphatic, heteroalicyclic,aryl, heteroaryl, alkylaryl, or alkylheteroaryl, wherein any of thealiphatic, alicyclic, heteroaliphatic, heteroalicyclic, alkylaryl, oralkylheteroaryl substituents described above and herein may besubstituted or unsubstituted, branched or unbranched, cyclic or acyclic,and wherein any of the aryl or heteroaryl substituents described aboveand herein may be substituted or unsubstituted. Additional examples ofgenerally applicable substituents are illustrated by the specificembodiments shown in the Examples that are described herein.

The term “heteroalicyclic”, as used herein, refers to compounds whichcombine the properties of heteroaliphatic and cyclic compounds andinclude but are not limited to saturated and unsaturated mono- orpolycyclic heterocycles such as morpholino, pyrrolidinyl, furanyl,thiofuranyl, pyrrolyl etc., which are optionally substituted with one ormore functional groups, as defined herein.

Additionally, it will be appreciated that any of the alicyclic orheteroalicyclic moieties described above and herein may comprise an arylor heteroaryl moiety fused thereto. Additional examples of generallyapplicable substituents are illustrated by the specific embodimentsshown in the Examples that are described herein.

The terms “halo” and “halogen” as used herein refer to an atom selectedfrom fluorine, chlorine, bromine and iodine.

The term “haloalkyl” denotes an alkyl group, as defined above, havingone, two, or three halogen atoms attached thereto and is exemplified bysuch groups as chloromethyl, bromoethyl, trifluoromethyl, and the like.

The term “heterocycloalkyl” or “heterocycle”, as used herein, refers toa non-aromatic 5-, 6- or 7-membered ring or a polycyclic group,including, but not limited to a bi- or tri-cyclic group comprising fusedsix-membered rings having between one and three heteroatomsindependently selected from oxygen, sulfur and nitrogen, wherein (i)each 5-membered ring has 0 to 1 double bonds and each 6-membered ringhas 0 to 2 double bonds, (ii) the nitrogen and sulfur heteroatoms may beoptionally be oxidized, (iii) the nitrogen heteroatom may optionally bequaternized, and (iv) any of the above heterocyclic rings may be fusedto an aryl or heteroaryl ring. Representative heterocycles include, butare not limited to, pyrrolidinyl, pyrazolinyl, pyrazolidinyl,imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl,isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, andtetrahydrofuryl. In certain embodiments, a “substituted heterocycloalkylor heterocycle” group is utilized and as used herein, refers to aheterocycloalkyl or heterocycle group, as defined above, substituted bythe independent replacement of one, two or three of the hydrogen atomsthereon with but are not limited to aliphatic; heteroaliphatic; aryl;heteroaryl; alkylaryl; alkylheteroaryl; alkoxy; aryloxy; heteroalkoxy;heteroaryloxy; alkylthio; arylthio; heteroalkylthio; heteroarylthio; F;Cl; Br; I; —OH; —NO₂; —CN; —CF₃; —CH₂CF₃; —CHCl₂; —CH₂OH; —CH₂CH₂OH;—CH₂NH₂; —CH₂SO₂CH₃; —C(O)R_(x); —CO₂(R_(x)); —CON(R_(x))₂; —OC(O)R_(x);—OCO₂R_(x); —OCON(R_(x))₂; —N(R_(x))₂; —S(O)₂R_(x); —NR—(CO)R_(x)wherein each occurrence of R_(x) independently includes, but is notlimited to, aliphatic, heteroaliphatic, aryl, heteroaryl, alkylaryl, oralkylheteroaryl, wherein any of the aliphatic, heteroaliphatic,alkylaryl, or alkylheteroaryl substituents described above and hereinmay be substituted or unsubstituted, branched or unbranched, cyclic oracyclic, and wherein any of the aryl or heteroaryl substitutentsdescribed above and herein may be substituted or unsubstituted.Additional examples or generally applicable substituents are illustratedby the specific embodiments shown in the Examples, which are describedherein.

As used herein, the terms “aliphatic”, “heteroaliphatic”, “alkyl”,“alkenyl”, “alkynyl”, “heteroalkyl”, “heteroalkenyl”, “heteroalkynyl”,and the like encompass substituted and unsubstituted, saturated andunsaturated, and linear and branched groups. Similarly, the terms“alicyclic”, “heteroalicyclic”, “heterocycloalkyl”, “heterocycle” andthe like encompass substituted and unsubstituted, and saturated andunsaturated groups. Additionally, the terms “cycloalkyl”,“cycloalkenyl”, “cycloalkynyl”, “heterocycloalkyl”,“heterocycloalkenyl”, “heterocycloalkynyl”, “aryl”, “heteroaryl” and thelike encompass both substituted and unsubstituted groups.

3) Synthetic Methodology

As described above, the present invention provides novel macrocycleshaving formula (I) a described above and in certain classes andsubclasses herein. An overview of an exemplary synthesis of theinventive compounds is provided below, as detailed in Schemes 1-7, andin the Exemplification herein. It will be appreciated that the methodsas described herein can be applied to each of the compounds as disclosedherein and equivalents thereof. Additionally, the reagents and startingmaterials are well known to those skilled in the art. Although thefollowing schemes describe certain exemplary compounds, it will beappreciated that the use of alternate starting materials will yieldother analogs of the invention. For example, compounds are describedbelow where X is O; however, it will be appreciated that alternatestarting materials and/or intermediates can be utilized to generatecompounds where X is NH, N-alkyl, CH₂, etc.

In general, compounds as provided herein especially modification where Yand Z together as CH═CH or CH₂CH₂, are prepared from assembly of thesethree segments in two different orders depending on position ofmodifications on the ring, as depicted below.

For R₄ modifications, a third route was use to incorporate R₄ asdepicted below:

For compounds with Y and Z are heteroatom such N, O or CO, a differentset of reaction condition was used to form these bonds in place of a C—Cbond formation. Certain analogs were prepared using variation of thesemethods shown above.

For R₉ analogs, compounds as provided herein, are prepared from ageneral advance intermediate in additional to the general methodsdescribed above, as depicted below (2):

In certain embodiments, this general advance intermediate can besynthesized from two components, an aromatic component, the synthesis ofwhich is depicted in Scheme 1 and is described in more detail inexamples herein, and a protected diol component, the synthesis of whichis depicted in Scheme 2 and is described in more detail in examplesherein. As depicted in Scheme 3, and as described in more detail inexamples herein, these two components are coupled, and subsequentreduction to generate the double bond occurs. Finally, macrocyclizationis effected to generate the macrolactone intermediate.

As depicted in Scheme 4, and as described in the examples herein, analternate route to the protected diol intermediate provides facileaccess to compounds where R₄ is halogen. Coupling of this intermediatewith the aromatic component described above and herein, providesadditional structures where R₄ is halogen, or, as depicted, F.

It will be appreciated that once the core intermediate structures areconstructed a variety of other analogues can be generated. In but oneexample, C14-O analogues are provided (R₉ as described herein). Forexample, Scheme 5 depicts the synthesis of these analogues using aMitsunobu reaction to functionalize the C14 hydroxyl moiety.

Alternatively, as depicted in Scheme 6, the hydroxyl functionality inthe advance intermediate can be replaced with an amine functionality.This amine can be further substituted (e.g., with methyl groups, asdepicted in Scheme 6) with a variety of functional groups as describedherein, using methods available to one of ordinary skill in the art.

Alternatively, as depicted in Schemes 7 and 9, the amine functionalitymay be introduced earlier in the synthesis. This amine can be furthersubstituted (e.g., with methyl or ethyl groups, as depicted in Schemes 7and 9) with a variety of functional groups as described herein, usingmethods available to one of ordinary skill in the art. A synthesis ofacyclic intermediate 20 is depicted in Scheme 8.

For special fused ring systems on the aromatic component, a differentaromatic segment is used in the place of the phenol. While synthesis ofthe aromatic fragment required special synthetic techniques, the overallflow remained, as depicted below (Scheme 10)

Synthetic Route to NF-2561

4) Research Uses, Formulation and Administration

According to the present invention, the inventive compounds may beassayed in any of the available assays known in the art for identifyingcompounds having antiangiogenic activity, anti-inflammatory activity,protein kinase inhibitory activity, NF-κB activation inhibitory activityactivity and AP-1 activation inhibitory activity. For example, the assaymay be cellular or non-cellular, in vivo or in vitro, high- orlow-throughput format, etc.

Thus, in one aspect, compounds of this invention which are of particularinterest include those which:

-   -   exhibit activity as inhibitors of NF-κB activation, AP-1        activation and protein kinases (e.g., MEKK1, MEK1, VEGFr,        PDGFr);    -   exhibit an antiproliferative or an antiangiogenic effect on        solid tumors;    -   exhibit an anti-inflammatory effect on suitable cell lines        maintained in vitro, or in animal studies using a scientifically        acceptable model;    -   are useful for the treatment of photoaging-related        disorders/conditions; and/or    -   exhibit a favorable therapeutic profile (e.g., safety, efficacy,        and stability).

As discussed above, certain of the compounds as described herein exhibitactivity generally as inhibitors of NF-κB activation, AP-1 activationand protein kinases. More specifically, compounds of the inventiondemonstrate immunosuppressive activity and thus the invention furtherprovides a method for treating an inflammatory disorder or autoimmunedisorders. Certain of the compounds as described herein also act asinhibitors of tumor growth and angiogenesis. The method involves theadministration of a therapeutically effective amount of the compound ora pharmaceutically acceptable derivative thereof to a subject(including, but not limited to a human or animal) in need of it. Incertain embodiments, the inventive compounds as useful for the treatmentof sepsis, glomerulonephropathy, rheumatoid arthritis (includingankylosing spondylitis), psoriatic arthritis, osteoarthritis,osteoporosis, allergic rhinitis, ocular inflammation, inflammatory boweldisease (crohn's disease and ulcerative colitis), multiple sclerosis,atopic dermatitis, psoriasis, asthma, inflammatory pulmonary disease,hepatitis, autoimmune disorders, systemic lupus erthematosus, allograftrejection/graft versus host disease, diabetes, AIDS, solid tumorcancers, leukemia, lymphomas, non-hodgkin's B-cell lymphomas, chronicallymphocytic leukemia (CLL), multiple myeloma, eczema, urticaria,myasthenia gravis, idiopathic thrombocytopenia purpura, cardiovasculardisease (e.g., myocardial infarction, atherosclerosis), hepatitis,glomerulonephropathy, productive nephritis, adenovirus,diseases/disorders of the central nervous system (e.g., stroke,Alzheimer's disease, epilepsy) and for the treatment of the symptoms ofmalaria, to name a few.

In certain other embodiments, compounds of the invention are useful forreducing photodamage, and thus, the invention further provides a methodfor treating photoaging-related disorders/conditions. In certainexemplary embodiments, compounds of the invention are useful for thetreatment and/or prevention of skin coarseness, wrinkling, mottledpigmentation, sallowness, laxity, telangiectasia, lentigines, purpuraand easy bruising, atrophy, fibrotic depigmented areas, and ultimatelypremalignant and malignant neoplasms. In certain other exemplaryembodiments, compounds of the invention are useful for the treatmentand/or prevention of wrinkles and/or skin cancer.

Pharmaceutical Compositions

As discussed above this invention provides novel compounds that havebiological properties useful for the treatment of inflammatory andautoimmune disorders, photoaging and cancer. The inventive compoundsalso find use in the prevention of restenosis of blood vessels subjectto traumas such as angioplasty and stenting. Accordingly, in anotheraspect of the present invention, pharmaceutical compositions areprovided, which comprise any one of the compounds described herein (or aprodrug, pharmaceutically acceptable salt or other pharmaceuticallyacceptable derivative thereof), and optionally comprise apharmaceutically acceptable carrier. In certain embodiments, thesecompositions optionally further comprise one or more additionaltherapeutic agents. Alternatively, a compound of this invention may beadministered to a patient in need thereof in combination with theadministration of one or more other therapeutic agents. For example,additional therapeutic agents for conjoint administration or inclusionin a pharmaceutical composition with a compound of this invention may bean immunomodulatory agent (e.g., an agent for the treatment of,rheumatoid arthritis, psoriasis, multiple sclerosis, or asthma) orantiangiogenesis agent or anticancer agent approved for the treatment ofcancer, as discussed in more detail herein, or it may be any one of anumber of agents undergoing approval in the Food and Drug Administrationthat ultimately obtain approval for the treatment of an immune disorderor cancer. It will also be appreciated that certain of the compounds ofpresent invention can exist in free form for treatment, or whereappropriate, as a pharmaceutically acceptable derivative thereof.According to the present invention, a pharmaceutically acceptablederivative includes, but is not limited to, pharmaceutically acceptablesalts, esters, salts of such esters, or a prodrug or other adduct orderivative of a compound of this invention which upon administration toa patient in need is capable of providing, directly or indirectly, acompound as otherwise described herein, or a metabolite or residuethereof.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts of amines, carboxylic acids, and other types ofcompounds, are well known in the art. For example, S. M. Berge, et al.describe pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 66: 1-19 (1977), incorporated herein byreference. The salts can be prepared in situ during the final isolationand purification of the compounds of the invention, or separately byreacting a free base or free acid function with a suitable reagent, asdescribed generally below. For example, a free base function can bereacted with a suitable acid. Furthermore, where the compounds of theinvention carry an acidic moiety, suitable pharmaceutically acceptablesalts thereof may, include metal salts such as alkali metal salts, e.g.sodium or potassium salts; and alkaline earth metal salts, e.g. calciumor magnesium salts. Examples of pharmaceutically acceptable, nontoxicacid addition salts are salts of an amino group formed with inorganicacids such as hydrochloric acid, hydrobromic acid, phosphoric acid,sulfuric acid and perchloric acid or with organic acids such as aceticacid, oxalic acid, maleic acid, tartaric acid, citric acid, succinicacid or malonic acid or by using other methods used in the art such asion exchange. Other pharmaceutically acceptable salts include adipate,alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate,borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hernisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Representative alkali or alkaline earth metal salts includesodium, lithium, potassium, calcium, magnesium, and the like. Furtherpharmaceutically acceptable salts include, when appropriate, nontoxicammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, loweralkyl sulfonate and aryl sulfonate.

Additionally, as used herein, the term “pharmaceutically acceptableester” refers to esters that hydrolyze in vivo and include those thatbreak down readily in the human body to leave the parent compound or asalt thereof. Suitable ester groups include, for example, those derivedfrom pharmaceutically acceptable aliphatic carboxylic acids,particularly alkanoic, alkenoic, cycloalkanoic and alkanedioic acids, inwhich each alkyl or alkenyl moeity advantageously has not more than 6carbon atoms. Examples of particular esters include formates, acetates,propionates, butyrates, acrylates and ethylsuccinates.

Furthermore, the term “pharmaceutically acceptable prodrugs” as usedherein refers to those prodrugs of the compounds of the presentinvention which are, within the scope of sound medical judgment,suitable for use in contact with the issues of humans and lower animalswith undue toxicity, irritation, allergic response, and the like,commensurate with a reasonable benefit/risk ratio, and effective fortheir intended use, as well as the zwitterionic forms, where possible,of the compounds of the invention. The term “prodrug” refers tocompounds that are rapidly transformed in vivo to yield the parentcompound of the above formula, for example by hydrolysis in blood. Athorough discussion is provided in T. Higuchi and V. Stella, Pro-drugsas Novel Delivery Systems, Vol. 14 of the A.C.S. Symposium Series, andin Edward B. Roche, ed., Bioreversible Carriers in Drug Design, AmericanPharmaceutical Association and Pergamon Press, 1987, both of which areincorporated herein by reference.

As described above, the pharmaceutical compositions of the presentinvention additionally comprise a pharmaceutically acceptable carrier,which, as used herein, includes any and all solvents, diluents, or otherliquid vehicle, dispersion or suspension aids, surface active agents,isotonic agents, thickening or emulsifying agents, preservatives, solidbinders, lubricants and the like, as suited to the particular dosageform desired. Remington's Pharmaceutical Sciences, Sixteenth Edition, E.W. Martin (Mack Publishing Co., Easton, Pa., 1980) discloses variouscarriers used in formulating pharmaceutical compositions and knowntechniques for the preparation thereof. Except insofar as anyconventional carrier medium is incompatible with the compounds of theinvention, such as by producing any undesirable biological effect orotherwise interacting in a deleterious manner with any othercomponent(s) of the pharmaceutical composition, its use is contemplatedto be within the scope of this invention. Some examples of materialswhich can serve as pharmaceutically acceptable carriers include, but arenot limited to, sugars such as lactose, glucose and sucrose; starchessuch as corn starch and potato starch; cellulose and its derivativessuch as sodium carboxymethyl cellulose, ethyl cellulose and celluloseacetate; powdered tragacanth; malt; gelatine; talc; excipients such ascocoa butter and suppository waxes; oils such as peanut oil, cottonseedoil; safflower oil, sesame oil; olive oil; corn oil and soybean oil;glycols; such as propylene glycol; esters such as ethyl oleate and ethyllaurate; agar; buffering agents such as magnesium hydroxide and aluminumhydroxide; alginic acid; pyrogenfree water; isotonic saline; Ringer'ssolution; ethyl alcohol, and phosphate buffer solutions, as well asother non-toxic compatible lubricants such as sodium lauryl sulfate andmagnesium stearate, as well as coloring agents, releasing agents,coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator.

Uses and Formulations of Compounds of the Invention

As described in more detail herein, in general, the present inventionprovides compounds useful for the treatment of inflammatory or immunedisorders and the treatment of cancer, particularly solid tumors.Without wishing to be bound by any particular theory, more generally,the compounds of the invention have been shown to inhibit NF-κB activityand the identification of NF-κB as a key player in the pathogenesis ofinflammation suggest that NF-κB targeted therapeutics may be effectivein inflammatory and immune disorders (see, generally, NF-κB in Defenseand Disease, J. Clin. Investig. 2001, 107, 7). Furthermore, certaincompounds of the invention have also been shown to inhibit receptortyrosine kinase activity such as VEGFr and PDGFr in vitro, as describedin more detail herein, and are useful for the treatment of cancer,including solid tumors (see, Angiogenesis: Potentials for PharmacologicIntervention in the Treatment of Cancer, Cardiovascular Diseases, andChronic Inflammation, Pharmacological Reviews, 2000, 52, 237).

As detailed in the exemplification herein, in assays to determine theability of compounds to inhibit NF-κB, certain inventive compoundsexhibited IC₅₀ values less than 10 μM. In certain other embodiments,inventive compounds exhibit IC₅₀ values less than 7.5 μM. In certainembodiments, inventive compounds exhibit IC₅₀ values less than 5 μM. Incertain other embodiments, inventive compounds exhibit IC₅₀ values lessthan 2.5 μM. In certain embodiments, inventive compounds exhibit IC₅₀values less than 1 μM. In certain embodiments, inventive compoundsexhibit IC₅₀ values less than 0.75 μM. In certain embodiments, inventivecompounds exhibit IC₅₀ values less than 0.5 μM. In certain embodiments,inventive compounds exhibit IC₅₀ values less than 0.25 μM. In certainembodiments, inventive compounds exhibit IC₅₀ values less than 0.1 μM.In certain other embodiments, inventive compounds exhibit IC₅₀ valuesless than 750 nM. In certain other embodiments, inventive compoundsexhibit IC₅₀ values less than 500 nM. In certain other embodiments,inventive compounds exhibit IC₅₀ values less than 250 nM. In certainother embodiments, inventive compounds exhibit IC₅₀ values less than 100nM. In other embodiments, exemplary compounds exhibited IC₅₀ values lessthan 75 nM. In other embodiments, exemplary compounds exhibited IC₅₀values less than 50 nM.

In still other embodiments, certain compounds were tested for theirability to inhibit the growth of tumor cell lines in vitro. Certain ofthese compounds exhibited IC₅₀ values less than 10 μM. In certain otherembodiments, inventive compounds exhibit IC₅₀ values less than 7.5 μM.In certain embodiments, inventive compounds exhibit IC₅₀ values lessthan 5 μM. In certain other embodiments, inventive compounds exhibitIC₅₀ values less than 2.5 μM. In certain embodiments, inventivecompounds exhibit IC₅₀ values less than 1 μM. In certain embodiments,inventive compounds exhibit IC₅₀ values less than 0.75 μM. In certainembodiments, inventive compounds exhibit IC₅₀ values less than 0.5 μM.In certain embodiments, inventive compounds exhibit IC₅₀ values lessthan 0.25 μM. In certain embodiments, inventive compounds exhibit IC₅₀values less than 0.1 μM. In certain other embodiments, inventivecompounds exhibit IC₅₀ values less than 750 nM. In certain otherembodiments, inventive compounds exhibit IC₅₀ values less than 500 nM.In certain other embodiments, inventive compounds exhibit IC₅₀ valuesless than 250 nM. In certain other embodiments, inventive compoundsexhibit IC₅₀ values less than 100 nM. In other embodiments, exemplarycompounds exhibited IC₅₀ values less than 75 nM. In other embodiments,exemplary compounds exhibited IC₅₀ values less than 50 nM.

As discussed above, compounds of the invention exhibit immunomodulatoryactivity and exhibit activity for the inhibition of angiogenesis throughinhibition of receptor tyrosine kinases. As such, the inventivecompounds as useful for the treatment of a variety of disorders,including, but not limited to, sepsis, glomerulonephropathy, rheumatoidarthritis (including ankylosing spondylitis), psoriatic arthritis,osteoarthritis, osteoporosis, allergic rhinitis, ocular inflammation,inflammatory bowel disease, atopic dermatitis, psoriasis, asthma,Crohn's disease, ulcerative colitis, inflammatory pulmonary disease,hepatitis, autoimmune disorders, diabetes, AIDS, solid tumor cancers,Leukemia, lymphomas, non-hodgkin's B-cell lymphomas, chronicallymphocytic leukemia (CLL), multiple myeloma, systemic lupuserythematosus, allograft rejection/graft versus host disease, eczema,uticaria, myasthenia gravis, idiopathic thrombocytopenia purpura,cardiovascular disease (e.g., myocardial infarction, atherosclerosis),hepatitis, productive nephritis, adenovirus, diseases/disorders of thecentral nervous system (stroke, Alzheimer's disease, epilepsy) and forthe treatment of the symptoms of malaria, to name a few. In certainembodiments, compounds of the invention are particularly useful for thetreatment of rheumatoid arthritis, psoriasis, multiple sclerosis, asthmaand cancer.

Rheumatoid Arthritis is a chronic syndrome characterized by nonspecific,usually symmetric inflammation of the peripheral joints, potentiallyresulting in progressive destruction of articular and periarticularstructures, with or without generalized manifestations (See, generally,The Merck Manual, 1999, Seventeenth Ed. Merck & Co., the entire contentsof which are hereby incorporated by reference). Studies in the pastestablished that presence of inflammatory cells and pro-inflammatorycytokines, such as TNFα, IL-1β are abundant in the diseased synovium.Increased macrophage-derived lining cells are prominent along with somelymphocytes and vascular changes in early disease. Although there is nota cure, reduction of circulatory pro-inflammatory cytokines (e.g. TNFα,IL-1β) through intervention of biological agents, such as Enbrel,Remicade or Anakinra demonstrated efficacy in reduction of symptoms andretarding the disease progression in clinical trials. Thus developing ofan agent such as described in this patent in modulation ofpro-inflammatory cytokines through NF-κB inhibition could bring greatbenefit to RA patients.

Psoriasis is a disorder for which there is no curative therapy, althoughin most cases acute attacks can be controlled. Psoriasis is a chronic,recurrent disease characterized by dry, well-circumscribed, silvery,scaling papules and plaques of various sizes, and has traditionally beenattributed to increased epidermal cell proliferation and concomitantdermal inflammation. The response of psoriasis to the immunosuppressivedrug cyclosporine suggests that the primary pathogenic factor may beimmunologic. Proliferation of epidermal cells has been also linked toAP-1 activation via stimulation from injury, radiation or stress to theskin (see, P. Angel et al., “Function and regulation of AP-1 subunits inskin physiology and pathology”, Oncogene, 2001, 20:2413-2423; and A.Grandjean-Laquerriere et al., “Relative contribution of NF-kB and AP-1in the modulation by Curcumin and pyrrolidine dithiocarbamate of theUVB-induced cytokine expression by keratinocytes”, Cytokine, 2002,18(3): 168-177, each of which is hereby incorporated by reference in itsentirety). Currently available treatment regimens for psoriasis includethe use of lubricants, keratolytics, topical cortisosteroids, sunlight,topical vitamin D derivatives, anthralin, and systemic antimetabolites(e.g., methotrexate), immunosuppressive drugs (e.g., cyclosporine,tacrolimus, mycophenolate, and mofetil). However, immunosuppressivedrugs are not yet approved for the treatment of psoriasis and otherdrugs, including corticosteriods, have severe side effects, includingexacerbations or pustular lesions (See, generally, The Merck Manual,1999, Seventeenth Ed. Merck & Co., the entire contents of which arehereby incorporated by reference). This invention is certainlyapplicable to this disease as well as a host of other related diseases,such as, psoriatic arthritis, ankylosing spondylitis, just to name afew.

Asthma is also believed to involve immunologic abnormalities andincreased inflammatory responses. Similarly to psoriasis, there is nocurative therapy. Thus the development of novel therapies such as this,preferably safe and curative, is desirable. This is also applied torelated immunologic disorders such as, graft rejection, SLE etc.

Angiogenesis, or the formation of new blood vessels out of pre-existingcapillaries, is a sequence of events that is fundamental to manyphysiologic and pathologic processes such as cancer, ischemic diseases,and chronical inflammation. With the identification of severalproangiogenic molecules such as vascular endothelial cell growth factor(VEGF), the fibroblast growth factors (FGFs) (see, Angiogenesis:Potentials for Pharmacologic Intervention in the Treatment of Cancer,Cardiovascular Diseases, and chronic Inflammation, PharmacologicalReviews, 2000, 52, 253). Thus, inhibition of receptor tyrosine kinase(such as VEGFr) activity has been subjects of various ongoing clinicaltrails. Certain compounds in this invention showed potent VEGFrinhibition. Thus, such application is expected.

As discussed above, the inventive compounds also find use in theprevention of restenosis of blood vessels subject to traumas such asangioplasty and stenting. For example, it is contemplated that thecompounds of the invention will be useful as a coating for implantedmedical devices, such as tubings, shunts, catheters, artificialimplants, pins, electrical implants such as pacemakers, and especiallyfor arterial or venous stents, including balloon-expandable stents. Incertain embodiments inventive compounds may be bound to an implantablemedical device, or alternatively, may be passively adsorbed to thesurface of the implantable device. In certain other embodiments, theinventive compounds may be formulated to be contained within, or,adapted to release by a surgical or medical device or implant, such as,for example, stents, sutures, indwelling catheters, prosthesis, and thelike.

In certain exemplary embodiments, the inventive compounds may be used ascoating for stents. A stent is typically an open tubular structure thathas a pattern (or patterns) of apertures extending from the outersurface of the stent to the lumen. It is commonplace to make stents ofbiocompatible metallic materials, with the patterns cut on the surfacewith a laser machine. The stent can be electro-polished to minimizesurface irregularities since these irregularities can trigger an adversebiological response. However, stents may still stimulate foreign bodyreactions that result in thrombosis or restenosis. To avoid thesecomplications, a variety of stent coatings and compositions have beenproposed in the prior art literature both to reduce the incidence ofthese complications or other complications and restore tissue functionby itself or by delivering therapeutic compound to the lumen. Forexample, drugs having antiproliferative and anti-inflammatory activitieshave been evaluated as stent coatings, and have shown promise inpreventing retenosis (See, for example, Presbitero P. et al., “Drugeluting stents do they make the difference?”, Minerva Cardioangiol,2002, 50(5):431-442; Ruygrok P. N. et al., “Rapamycin in cardiovascularmedicine”, Intern. Med. J., 2003, 33(3):103-109; and Marx S. O. et al.,“Bench to bedside: the development of rapamycin and its application tostent restenosis”, Circulation, 2001, 104(8):852-855, each of thesereferences is incorporated herein by reference in its entirety).Accordingly, without wishing to be bound to any particular theory,Applicant proposes that inventive compounds having anti-inflammatoryand/or antiproliferative effects can be used as stent coatings and/or instent drug delivery devices, inter alia for the prevention of restenosisor reduction of restenosis rate. A variety of compositions and methodsrelated to stent coating and/or local stent drug delivery for preventingrestenosis are known in the art (see, for example, U.S. Pat. Nos.6,517,889; 6,273,913; 6,258,121; 6,251,136; 6,248,127; 6,231,600;6,203,551; 6,153,252; 6,071,305; 5,891,507; 5,837,313 and published U.S.patent application No.: US2001/0027340, each of which is incorporatedherein by reference in its entirety). For example, stents may be coatedwith polymer-drug conjugates by dipping the stent in polymer-drugsolution or spraying the stent with such a solution. In certainembodiment, suitable materials for the implantable device includebiocompatible and nontoxic materials, and may be chosen from the metalssuch as nickel-titanium alloys, steel, or biocompatible polymers,hydrogels, polyurethanes, polyethylenes, ethylenevinyl acetatecopolymers, etc. In certain embodiments, the inventive compound, iscoated onto a stent for insertion into an artery or vein followingballoon angioplasty.

The invention may be described therefore, in certain broad aspects as amethod of inhibiting arterial restenosis or arterial occlusion followingvascular trauma comprising administering to a subject in need thereof, acomposition comprising an inventive compound conjugated to a suitablepolymer or polymeric material. In the practice of the method, thesubject may be a coronary bypass, vascular surgery, organ transplant orcoronary or any other arterial angioplasty patient, for example, and thecomposition may be administered directly, intravenously, or even coatedon a stent to be implanted at the sight of vascular trauma.

In another aspect, the invention encompasses implants and surgical ormedical devices, including stents and grafts, coated with or otherwiseconstructed to contain and/or release any of the inventive compoundsdisclosed herein. In certain embodiments, the compounds haveanti-inflammatory and/or antiproliferative activities. In certain otherembodiments, the compounds inhibit smooth muscle cell proliferation.Representative examples of the inventive implants and surgical ormedical devices include cardiovascular devices (e.g., implantable venouscatheters, venous ports, tunneled venous catheters, chronic infusionlines or ports, including hepatic artery infusion catheters, pacemakerwires, implantable defibrillators); neurologic/neurosurgical devices(e.g., ventricular peritoneal shunts, ventricular atrial shunts, nervestimulator devices, dural patches and implants to prevent epiduralfibrosis post-laminectomy, devices for continuous subarachnoidinfusions); gastrointestinal devices (e.g., chronic indwellingcatheters, feeding tubes, portosystemic shunts, shunts for ascites,peritoneal implants for drug delivery, peritoneal dialysis catheters,implantable meshes for hernias, suspensions or solid implants to preventsurgical adhesions, including meshes); genitourinary devices (e.g.,uterine implants, including intrauterine devices (IUDs) and devices toprevent endometrial hyperplasia, fallopian tubal implants, includingreversible sterilization devices, fallopian tubal stents, artificialsphincters and periurethral implants for incontinence, ureteric stents,chronic indwelling catheters, bladder augmentations, or wraps or splintsfor vasovasostomy); phthalmologic implants (e.g., multino implants andother implants for neovascular glaucoma, drug eluting contact lenses forpterygiums, splints for failed dacrocystalrhinostomy, drug elutingcontact lenses for corneal neovascularity, implants for diabeticretinopathy, drug eluting contact lenses for high risk cornealtransplants); otolaryngology devices (e.g., ossicular implants,Eustachian tube splints or stents for glue ear or chronic otitis as analternative to transtempanic drains); plastic surgery implants (e.g.,prevention of fibrous contracture in response to gel- orsaline-containing breast implants in the subpectoral or subglandularapproaches or post-mastectomy, or chin implants), and orthopedicimplants (e.g., cemented orthopedic prostheses).

Implants and other surgical or medical devices may be coated with (orotherwise adapted to release) compositions of the present invention in avariety of manners, including for example: (a) by directly affixing tothe implant or device an inventive compound or composition (e.g., byeither spraying the implant or device with a polymer/drug film, or bydipping the implant or device into a polymer/drug solution, or by othercovalent or noncovalent means); (b) by coating the implant or devicewith a substance such as a hydrogel which will in turn absorb theinventive compound or composition; (c) by interweaving inventivecompound- or composition-coated thread (or the polymer itself formedinto a thread) into the implant or device; (d) by inserting the implantor device into a sleeve or mesh which is comprised of or coated with aninventive compound or composition; (e) constructing the implant ordevice itself with an inventive compound or composition; or (f) byotherwise adapting the implant or device to release the inventivecompound. In certain embodiments, the composition should firmly adhereto the implant or device during storage and at the time of insertion.The inventive compound or composition should also preferably not degradeduring storage, prior to insertion, or when warmed to body temperatureafter insertion inside the body (if this is required). In addition, itshould preferably coat the implant or device smoothly and evenly, with auniform distribution of inventive compound, while not changing the stentcontour. Within preferred embodiments of the invention, the inventiveimplant or device should provide a uniform, predictable, prolongedrelease of the inventive compound or composition into the tissuesurrounding the implant or device once it has been deployed. Forvascular stents, in addition to the above properties, the compositionshould not render the stent thrombogenic (causing blood clots to form),or cause significant turbulence in blood flow (more than the stentitself would be expected to cause if it was uncoated).

In the case of stents, a wide variety of stents may be developed tocontain and/or release the inventive compounds or compositions providedherein, including esophageal stents, gastrointestinal stents, vascularstents, biliary stents, colonic stents, pancreatic stents, ureteric andurethral stents, lacrimal stents, Eustachian tube stents, fallopian tubestents and tracheal/bronchial stents (See, for example, U.S. Pat. No.6,515,016, the entire contents of which are incorporated herein byreference). Stents may be readily obtained from commercial sources, orconstructed in accordance with well-known techniques. Representativeexamples of stents include those described in U.S. Pat. No. 4,768,523,entitled “Hydrogel Adhesive”; U.S. Pat. No. 4,776,337, entitled“Expandable Intraluminal Graft, and Method and Apparatus for Implantingand Expandable Intraluminal Graft”; U.S. Pat. No. 5,041,126 entitled“Endovascular Stent and Delivery System”; U.S. Pat. No. 5,052,998entitled “Indwelling Stent and Method of Use”; U.S. Pat. No. 5,064,435entitled “Self-Expanding Prosthesis Having Stable Axial Length”; U.S.Pat. No. 5,089,606, entitled “Water-insoluble Polysaccharide HydrogelFoam for Medical Applications”; U.S. Pat. No. 5,147,370, entitled“Nitinol Stent for Hollow Body Conduits”; U.S. Pat. No. 5,176,626,entitled “Indwelling Stent”; U.S. Pat. No. 5,213,580, entitled“Biodegradable Polymeric Endoluminal Sealing Process”; and U.S. Pat. No.5,328,471, entitled “Method and Apparatus for Treatment of Focal Diseasein Hollow Tubular Organs and Other Tissue Lumens.”

As discussed above, the stent coated with (or otherwise adapted torelease) compositions of the present invention may be used to eliminatea vascular obstruction and prevent restenosis or reduce the rate ofrestenosis. Within other aspects of the present invention, stents coatedwith (or otherwise adapted to release) compositions of the presentinvention are provided for expanding the lumen of a body passageway.Specifically, a stent having a generally tubular structure, and asurface coated with (or otherwise adapted to release) an inventivecompound or composition may be inserted into the passageway, such thatthe passageway is expanded. In certain embodiments, the stent coatedwith (or otherwise adapted to release) compositions of the presentinvention may be used to eliminate a biliary, gastrointestinal,esophageal, tracheal/bronchial, urethral or vascular obstruction.

In another aspect of the invention, methods for the treatment of immunedisorders and cancer are provided comprising administering atherapeutically effective amount of a compound of formula (I), asdescribed herein, to a subject in need thereof. In certain embodiments,the inventive compounds are useful for the treatment of rheumatoidarthritis, psoriasis, multiple sclerosis, asthma and cancer. It will beappreciated that the compounds and compositions, according to the methodof the present invention, may be administered using any amount and anyroute of administration effective for the treatment of inflammatorydisorders, including but not limited to rheumatoid arthritis, psoriasis,multiple sclerosis, asthma and cancer. Thus, the expression “effectiveamount” as used herein, refers to a sufficient amount of agent toinhibit the growth of tumor cells, or refers to a sufficient amount toreduce the effects of rheumatoid arthritis, psoriasis, asthma andcancer, (or any inflammatory response or disorder). The exact amountrequired will vary from subject to subject, depending on the species,age, and general condition of the subject, the severity of the diseases,the particular anticancer agent, its mode of administration, and thelike. The compounds of the invention are preferably formulated in dosageunit form for ease of administration and uniformity of dosage. Theexpression “dosage unit form” as used herein refers to a physicallydiscrete unit of therapeutic agent appropriate for the patient to betreated. It will be understood, however, that the total daily usage ofthe compounds and compositions of the present invention will be decidedby the attending physician within the scope of sound medical judgment.The specific therapeutically effective dose level for any particularpatient or organism will depend upon a variety of factors including thedisorder being treated and the severity of the disorder; the activity ofthe specific compound employed; the specific composition employed; theage, body weight, general health, sex and diet of the patient; the timeof administration, route of administration, and rate of excretion of thespecific compound employed; the duration of the treatment; drugs used incombination or coincidental with the specific compound employed; andlike factors well known in the medical arts (see, for example, Goodmanand Gilman's, “The Pharmacological Basis of Therapeutics”, TenthEdition, A. Gilman, J. Hardman and L. Limbird, eds., McGraw-Hill Press,155-173, 2001, which is incorporated herein by reference in itsentirety).

In certain other embodiments, methods are provided for using theinventive implants and other surgical or medical devices coated with (orotherwise adapted to release) compounds and compositions of the presentinvention. In certain embodiments, methods are provided for preventingrestenosis, comprising inserting a stent into an obstructed bloodvessel, the stent having a generally tubular structure, the surface ofthe structure being coated with (or otherwise adapted to release) aninventive compound or composition, such that the obstruction iseliminated and the inventive compound or composition is delivered inamounts effective to prevent restenosis. In other embodiments, methodsare provided for preventing restenosis, comprising inserting a stentinto an obstructed blood vessel, the stent having a generally tubularstructure, the surface of the structure being coated with (or otherwiseadapted to release) an inventive compound or composition, such that theobstruction is eliminated and the inventive compound or composition isdelivered in amounts effective to inhibit smooth muscle cellproliferation.

Within other aspects of the present invention, methods are provided forexpanding the lumen of a body passageway, comprising inserting a stentinto the passageway, the stent having a generally tubular structure, thesurface of the structure being coated with (or otherwise adapted torelease) an inventive compound or composition, such that the passagewayis expanded. In certain embodiments, the lumen of a body passageway isexpanded in order to eliminate a biliary, gastrointestinal, esophageal,tracheal/bronchial, urethral and/or vascular obstruction.

In certain embodiments, methods are provided for eliminating biliaryobstructions, comprising inserting a biliary stent into a biliarypassageway, the stent having a generally tubular structure, the surfaceof the structure being coated with (or otherwise adapted to release) aninventive compound or composition, such that the biliary obstruction iseliminated. Briefly, tumor overgrowth of the common bile duct results inprogressive cholestatic jaundice which is incompatible with life.Generally, the biliary system which drains bile from the liver into theduodenum is most often obstructed by (1) a tumor composed of bile ductcells (cholangiocarcinoma), (2) a tumor which invades the bile duct(e.g., pancreatic carcinoma), or (3) a tumor which exerts extrinsicpressure and compresses the bile duct (e.g., enlarged lymph nodes). Bothprimary biliary tumors, as well as other tumors which cause compressionof the biliary tree may be treated utilizing stents Implants and othersurgical or medical devices may be coated with (or otherwise adapted torelease) compositions of the present invention. One example of primarybiliary tumors are adenocarcinomas (which are also called Klatskintumors when found at the bifurcation of the common hepatic duct). Thesetumors are also referred to as biliary carcinomas,choledocholangiocarcinomas, or adenocarcinomas of the biliary system.Benign tumors which affect the bile duct (e.g., adenoma of the biliarysystem), and, in rare cases, squamous cell carcinomas of the bile ductand adenocarcinomas of the gallbladder, may also cause compression ofthe biliary tree and therefore, result in biliary obstruction.Compression of the biliary tree is most commonly due to tumors of theliver and pancreas which compress and therefore obstruct the ducts. Mostof the tumors from the pancreas arise from cells of the pancreaticducts. This is a highly fatal form of cancer (5% of all cancer deaths;26,000 new cases per year in the U.S.) with an average of 6 monthssurvival and a 1 year survival rate of only 10%. When these tumors arelocated in the head of the pancreas they frequently cause biliaryobstruction, and this detracts significantly from the quality of life ofthe patient. While all types of pancreatic tumors are generally referredto as “carcinoma of the pancreas” there are histologic subtypesincluding: adenocarcinoma, adenosquamous carcinoma, cystadenocarcinoma,and acinar cell carcinoma. Hepatic tumors, as discussed above, may alsocause compression of the biliary tree, and therefore cause obstructionof the biliary ducts.

In certain embodiments, a biliary stent is first inserted into a biliarypassageway in one of several ways: from the top end by inserting aneedle through the abdominal wall and through the liver (a percutaneoustranshepatic cholangiogram or “PTC”); from the bottom end by cannulatingthe bile duct through an endoscope inserted through the mouth, stomach,and duodenum (an endoscopic retrograde cholangiogram or “ERCP”); or bydirect incision during a surgical procedure. In certain embodiments, apreinsertion examination, PTC, ERCP, or direct visualization at the timeof surgery is performed to determine the appropriate position for stentinsertion. A guidewire is then advanced through the lesion, and overthis a delivery catheter is passed to allow the stent to be inserted inits collapsed form. If the diagnostic exam was a PTC, the guidewire anddelivery catheter is inserted via the abdominal wall, while if theoriginal exam was an ERCP the stent may be placed via the mouth. Thestent is then positioned under radiologic, endoscopic, or direct visualcontrol taking particular care to place it precisely across thenarrowing in the bile duct. The delivery catheter is then removedleaving the stent standing as a scaffolding which holds the bile ductopen. A further cholangiogram may be performed to document that thestent is appropriately positioned.

In certain embodiments, methods are provided for eliminating esophagealobstructions, comprising inserting an esophageal stent into anesophagus, the stent having a generally tubular structure, the surfaceof the structure being coated with (or otherwise adapted to release) aninventive compound or composition, such that the esophageal obstructionis eliminated. Briefly, the esophagus is the hollow tube whichtransports food and liquids from the mouth to the stomach. Cancer of theesophagus or invasion by cancer arising in adjacent organs (e.g., cancerof the stomach or lung) results in the inability to swallow food orsaliva. In certain embodiments, a preinsertion examination, usually abarium swallow or endoscopy is performed in order to determine theappropriate position for stent insertion. A catheter or endoscope maythen be positioned through the mouth, and a guidewire is advancedthrough the blockage. A stent delivery catheter is passed over theguidewire under radiologic or endoscopic control, and a stent is placedprecisely across the narrowing in the esophagus. A post-insertionexamination, usually a barium swallow x-ray, may be utilized to confirmappropriate positioning.

In certain embodiments, methods are provided for eliminating colonicobstructions, comprising inserting a colonic stent into a colon, thestent having a generally tubular structure, the surface of the structurebeing coated with (or otherwise adapted to release) an inventivecompound or composition, such that the colonic obstruction iseliminated. Briefly, the colon is the hollow tube which transportsdigested food and waste materials from the small intestines to the anus.Cancer of the rectum and/or colon or invasion by cancer arising inadjacent organs (e.g., cancer of the uterus, ovary, bladder) results inthe inability to eliminate feces from the bowel. In certain embodiments,a preinsertion examination, usually a barium enema or colonoscopy isperformed in order to determine the appropriate position for stentinsertion. A catheter or endoscope may then be positioned through theanus, and a guidewire is advanced through the blockage. A stent deliverycatheter is passed over the guidewire under radiologic or endoscopiccontrol, and a stent is placed precisely across the narrowing in thecolon or rectum. A post-insertion examination, usually a barium enemax-ray, may be utilized to confirm appropriate positioning.

In certain embodiments, methods are provided for eliminatingtracheal/bronchial obstructions, comprising inserting atracheal/bronchial stent into a trachea or bronchi, the stent having agenerally tubular structure, the surface of the structure being coatedwith (or otherwise adapted to release) an inventive compound orcomposition, such that the tracheal/bronchial obstruction is eliminated.Briefly, the trachea and bronchi are tubes which carry air from themouth and nose to the lungs. Blockage of the trachea by cancer, invasionby cancer arising in adjacent organs (e.g., cancer of the lung), orcollapse of the trachea or bronchi due to chondromalacia (weakening ofthe cartilage rings) results in inability to breathe. In certainembodiments, preinsertion examination, usually an endoscopy, isperformed in order to determine the appropriate position for stentinsertion. A catheter or endoscope is then positioned through the mouth,and a guidewire advanced through the blockage. A delivery catheter isthen passed over the guidewire in order to allow a collapsed stent to beinserted. The stent is placed under radiologic or endoscopic control inorder to place it precisely across the narrowing. The delivery cathetermay then be removed leaving the stent standing as a scaffold on its own.A post-insertion examination, usually a bronchoscopy may be utilized toconfirm appropriate positioning.

In certain embodiments, methods are provided for eliminating urethralobstructions, comprising inserting a urethral stent into a urethra, thestent having a generally tubular structure, the surface of the structurebeing coated with (or otherwise adapted to release) an inventivecompound or composition, such that the urethral obstruction iseliminated. Briefly, the urethra is the tube which drains the bladderthrough the penis. Extrinsic narrowing of the urethra as it passesthrough the prostate, due to hypertrophy of the prostate, occurs invirtually every man over the age of 60 and causes progressive difficultywith urination. In certain embodiments, a preinsertion examination,usually an endoscopy or urethrogram is first performed in order todetermine the appropriate position for stent insertion, which is abovethe external urinary sphincter at the lower end, and close to flush withthe bladder neck at the upper end. An endoscope or catheter is thenpositioned through the penile opening and a guidewire advanced into thebladder. A delivery catheter is then passed over the guidewire in orderto allow stent insertion. The delivery catheter is then removed, and thestent expanded into place. A post-insertion examination, usuallyendoscopy or retrograde urethrogram, may be utilized to confirmappropriate position.

In certain embodiments, methods are provided for eliminating vascularobstructions, comprising inserting a vascular stent into a blood vessel,the stent having a generally tubular structure, the surface of thestructure being coated with (or otherwise adapted to release) aninventive compound or composition, such that the vascular obstruction iseliminated. Briefly, stents may be placed in a wide array of bloodvessels, both arteries and veins, to prevent recurrent stenosis at thesite of failed angioplasties, to treat narrowings that would likely failif treated with angioplasty, and to treat post-surgical narrowings(e.g., dialysis graft stenosis). Suitable sites include, but are notlimited to, the iliac, renal, and coronary arteries, the superior venacava, and in dialysis grafts. In certain embodiments, angiography isfirst performed in order to localize the site for placement of thestent. This is typically accomplished by injecting radiopaque contrastthrough a catheter inserted into an artery or vein as an x-ray is taken.A catheter may then be inserted either percutaneously or by surgery intothe femoral artery, brachial artery, femoral vein, or brachial vein, andadvanced into the appropriate blood vessel by steering it through thevascular system under fluoroscopic guidance. A stent may then bepositioned across the vascular stenosis. A post-insertion angiogram mayalso be utilized in order to confirm appropriate positioning.

In certain other embodiments, compounds of the invention are useful forreducing photodamage, and thus, the invention further provides a methodfor treating photoaging-related disorders/conditions. Photoaging is aterm used to describe the changes in appearance and function of skin asa result of repeated exposure to sunlight. The ultraviolet (UV)component of sunlight, particularly middle UV (called UVB, 290-320 nmwavelength) is the principal causative agent of photoaging. The extentof UVB exposure required to cause photoaging is not currently known.Repeated exposure to UVB at levels that cause erythema and tanning are,however, commonly associated with photoaging. Clinically, photoaging ischaracterized by coarseness, wrinkling, mottled pigmentation,sallowness, laxity, telangiectasia, lentigines, purpura and easybruising, atrophy, fibrotic depigmented areas, and ultimatelypremalignant and malignant neoplasms. Photoaging commonly occurs in skinthat is habitually exposed to sunlight such as the face, ears, baldareas of the scalp, neck, and hands.

Procedures for preventing photoaging of unaged skin and treating alreadyphotoaged skin are available. Sunscreens are commonly used to preventphotoaging of skin areas that are habitually exposed to sunlight.Sunscreens are topical preparations that absorb, reflect or scatter UV.Some are based on opaque particulate materials such as zinc oxide,titanium oxide, clays and ferric chloride. Because such preparations arevisible and occlusive many people consider these opaque formulationscosmetically unacceptable. Other sunscreens contain chemicals such ap-aminobenzoic acid (PABA), oxybenzone, dioxybenzone, ethylhexyl-methoxycinnamide and butylmethoxydibenzoylmethane that are nonopaque andcolorless because they do not absorb light of visible wavelengths. Whilethese nonopaque sunscreens may be more acceptable cosmetically they arestill relatively short-lived and susceptible to being removed by washingor perspiration. Additionally all sunscreens reduce vitamin Dproduction.

It is known that transcription factors AP-1 and NF-κB are activated inmammalian cells exposed to UV light. It has also been shown thatinhibition of MAP kinase/ERK kinase 1 (MEK-1) significantly inhibitedUVB induced ERK activation (See, Chen et al., “Activation of p38 MAPkinase and ERK are required for ultraviolet-B induced c-fos geneexpression in human keratinocytes”, Oncogene, 18:7469-7476, 1999; theentire contents of which are incorporated herein by reference).Accordingly, without wishing to be bound to any particular theory,Applicant proposes that the compounds of the invention may find use inthe treatment of skin damages caused by UVB exposure. For additionalreferences on MAP kinases and photoaging, see Li et al., “Rays andarrays: the transcriptional program in the response of human epidermalkeratotinocutes to UVB illumination”, The FASEB Journal express article10.1096/fj.01-01172fje, published online Sep. 17, 2001; U.S. Pat. No.5,837,224 and U.S. Patent Application No.: 20020106339, each of which ishereby incorporated by reference in its entirety.

Thus the invention provides compositions for preventing or treatingUVB-induced photodamage comprising an inventive compound; and apharmaceutically acceptable carrier. In certain embodiments, theinventive compound is present in an amount effective to inhibit Map/Erkkinase. In certain other embodiments, the inventive compositions furthercomprise a cosmetic ingredient. In certain exemplary embodiments, thecosmetic ingredient is a fragrance. In certain other exemplaryembodiments, the cosmetic ingredient is a sunscreen. In certainembodiments, the inventive compositions exist as pharmaceuticallyacceptable topical formulations.

The present invention additionally encompasses methods of providingprotection against long-term UVB induced photodamage to a subject, saidmethod comprising: administering to the subject in need thereof acomposition comprising an inventive compound; and a pharmaceuticallyacceptable carrier or diluent. In certain embodiments, the compositionis administered topically. The present invention additionallyencompasses methods of providing protection against long-term UVBinduced photodamage to a subject, said method comprising: providing thesubject with a composition comprising an inventive compound; andproviding the subject with instructions for using said composition toprevent photodamage. In certain embodiments, the composition isformulated so that it may be administered topically. In certainembodiments, the inventive compound is present in an amount effective toinhibit Map/Erk kinase. In certain embodiments, the instructionscomprise directions to apply the composition to the skin prior to sunexposure. In certain exemplary embodiments, the composition furthercomprises a cosmetic ingredient. In certain exemplary embodiments, thecosmetic ingredient is a fragrance. In certain other exemplaryembodiments, the cosmetic ingredient is a sunscreen. In certainembodiment, a method is provided for treating and/or preventing skincoarseness, wrinkling, mottled pigmentation, sallowness, laxity,telangiectasia, lentigines, purpura and easy bruising, atrophy, fibroticdepigmented areas, and ultimately premalignant and malignant neoplasms.In certain exemplary embodiments, the present invention provides amethod for treating and/or preventing wrinkles and/or skin cancer.

In certain embodiments, the present invention provides kits forpreventing long-term UVB induced photodamage in a subject, said kitcomprising: a composition comprising an inventive compound; andinstructions for using the composition to prevent photodamage. Incertain embodiments, the composition is formulated for topicaladministration. In certain embodiments, the inventive compound ispresent in an amount effective to inhibit Map/Erk kinase. In certainembodiments, the instructions comprise directions to apply thecomposition to the skin prior to sun exposure. In certain exemplaryembodiments, the composition further comprises a cosmetic ingredient. Incertain exemplary embodiments, the cosmetic ingredient is a fragrance.In certain other exemplary embodiments, the cosmetic ingredient is asunscreen.

Furthermore, after formulation with an appropriate pharmaceuticallyacceptable carrier in a desired dosage, the pharmaceutical compositionsof this invention can be administered to humans and other animalsorally, rectally, parenterally, intracisternally, intravaginally,intraperitoneally, topically (as by powders, ointments, creams ordrops), bucally, as an oral or nasal spray, or the like, depending onthe severity of the infection being treated. In certain embodiments, thecompounds of the invention may be administered at dosage levels of about0.001 mg/kg to about 50 mg/kg, from about 0.01 mg/kg to about 25 mg/kg,or from about 0.1 mg/kg to about 10 mg/kg of subject body weight perday, one or more times a day, to obtain the desired therapeutic effect.It will also be appreciated that dosages smaller than 0.001 mg/kg orgreater than 50 mg/kg (for example 50-100 mg/kg) can be administered toa subject. In certain embodiments, compounds are administered orally orparenterally.

Liquid dosage forms for oral administration include, but are not limitedto, pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active compounds,the liquid dosage forms may contain inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fattyacid esters of sorbitan, and mixtures thereof. Besides inert diluents,the oral compositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This may be accomplished by the use of a liquid suspension orcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolutionthat, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle. Injectable depot forms are made by forming microencapsulematrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude (poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions, which are compatible with body tissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like. The solid dosage forms of tablets, dragees, capsules, pills,and granules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes. Solid compositions of a similartype may also be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polyethylene glycols and the like.

The active compounds can also be in microencapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose and starch. Such dosage forms may alsocomprise, as in normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such asmagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositions,which can be used, include polymeric substances and waxes.

The present invention encompasses pharmaceutically acceptable topicalformulations of inventive compounds. The term “pharmaceuticallyacceptable topical formulation”, as used herein, means any formulationwhich is pharmaceutically acceptable for intradermal administration of acompound of the invention by application of the formulation to theepidermis. In certain embodiments of the invention, the topicalformulation comprises a carrier system. Pharmaceutically effectivecarriers include, but are not limited to, solvents (e.g., alcohols, polyalcohols, water), creams, lotions, ointments, oils, plasters, liposomes,powders, emulsions, microemulsions, and buffered solutions (e.g.,hypotonic or buffered saline) or any other carrier known in the art fortopically administering pharmaceuticals. A more complete listing ofart-known carriers is provided by reference texts that are standard inthe art, for example, Remington's Pharmaceutical Sciences, 16th Edition,1980 and 17th Edition, 1985, both published by Mack Publishing Company,Easton, Pa., the disclosures of which are incorporated herein byreference in their entireties. In certain other embodiments, the topicalformulations of the invention may comprise excipients. Anypharmaceutically acceptable excipient known in the art may be used toprepare the inventive pharmaceutically acceptable topical formulations.Examples of excipients that can be included in the topical formulationsof the invention include, but are not limited to, preservatives,antioxidants, moisturizers, emollients, buffering agents, solubilizingagents, other penetration agents, skin protectants, surfactants, andpropellants, and/or additional therapeutic agents used in combination tothe inventive compound. Suitable preservatives include, but are notlimited to, alcohols, quaternary amines, organic acids, parabens, andphenols. Suitable antioxidants include, but are not limited to, ascorbicacid and its esters, sodium bisulfite, butylated hydroxytoluene,butylated hydroxyanisole, tocopherols, and chelating agents like EDTAand citric acid. Suitable moisturizers include, but are not limited to,glycerine, sorbitol, polyethylene glycols, urea, and propylene glycol.Suitable buffering agents for use with the invention include, but arenot limited to, citric, hydrochloric, and lactic acid buffers. Suitablesolubilizing agents include, but are not limited to, quaternary ammoniumchlorides, cyclodextrins, benzyl benzoate, lecithin, and polysorbates.Suitable skin protectants that can be used in the topical formulationsof the invention include, but are not limited to, vitamin E oil,allatoin, dimethicone, glycerin, petrolatum, and zinc oxide.

In certain embodiments, the pharmaceutically acceptable topicalformulations of the invention comprise at least a compound of theinvention and a penetration enhancing agent. The choice of topicalformulation will depend or several factors, including the condition tobe treated, the physicochemical characteristics of the inventivecompound and other excipients present, their stability in theformulation, available manufacturing equipment, and costs constraints.As used herein the term “penetration enhancing agent” means an agentcapable of transporting a pharmacologically active compound through thestratum corneum and into the epidermis or dermis, preferably, withlittle or no systemic absorption. A wide variety of compounds have beenevaluated as to their effectiveness in enhancing the rate of penetrationof drugs through the skin. See, for example, Percutaneous PenetrationEnhancers, Maibach H. I. and Smith H. E. (eds.), CRC Press, Inc., BocaRaton, Fla. (1995), which surveys the use and testing of various skinpenetration enhancers, and Buyuktimkin et al., Chemical Means ofTransdermal Drug Permeation Enhancement in Transdermal and Topical DrugDelivery Systems, Gosh T. K., Pfister W. R., Yum S. I. (Eds.),Interpharm Press Inc., Buffalo Grove, Ill. (1997). In certain exemplaryembodiments, penetration agents for use with the invention include, butare not limited to, triglycerides (e.g., soybean oil), aloe compositions(e.g., aloe-vera gel), ethyl alcohol, isopropyl alcohol,octolyphenylpolyethylene glycol, oleic acid, polyethylene glycol 400,propylene glycol, N-decylmethylsulfoxide, fatty acid esters (e.g.,isopropyl myristate, methyl laurate, glycerol monooleate, and propyleneglycol monooleate) and N-methylpyrrolidone.

In certain embodiments, the compositions may be in the form ofointments, pastes, creams, lotions, gels, powders, solutions, sprays,inhalants or patches. In certain exemplary embodiments, formulations ofthe compositions according to the invention are creams, which mayfurther contain saturated or unsaturated fatty acids such as stearicacid, palmitic acid, oleic acid, palmito-oleic acid, cetyl or oleylalcohols, stearic acid being particularly preferred. Creams of theinvention may also contain a non-ionic surfactant, for example,polyoxy-40-stearate. In certain embodiments, the active component isadmixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, eardrops, and eye drops are also contemplated asbeing within the scope of this invention. Additionally, the presentinvention contemplates the use of transdermal patches, which have theadded advantage of providing controlled delivery of a compound to thebody. Such dosage forms are made by dissolving or dispensing thecompound in the proper medium. As discussed above, penetration enhancingagents can also be used to increase the flux of the compound across theskin. The rate can be controlled by either providing a rate controllingmembrane or by dispersing the compound in a polymer matrix or gel.

In certain embodiments, after application of the topical formulation tothe epidermis, the area may be covered with a dressing. The term“dressing”, as used herein, means a covering designed to protect atopically applied drug formulation. “Dressing” includes coverings suchas a bandage, which may be porous or non-porous and various inertcoverings, e.g., a plastic film wrap or other non-absorbent film. Theterm “dressing” also encompasses non-woven or woven coverings,particularly elastomeric coverings, which allow for heat and vaportransport. These dressings allow for cooling of the treated area, whichprovides for greater comfort.

In certain exemplary embodiments, pharmaceutically acceptable topicalformulations of the invention are contained in a patch that is appliedadjacent to the area of skin to be treated. As used herein a “patch”comprises at least a topical formulation and a covering layer, suchthat, the patch can be placed over the area of skin to be treated.Preferably, but not necessarily, the patch is designed to maximize drugdelivery through the stratum corneum and into the epidermis or dermis,reduce lag time, promote uniform absorption, and/or reduce mechanicalrub-off. In certain embodiments, when the intended use comprises thetreatment of a skin condition (e.g., psoriasis), the patch is designedto minimize absorption into the circulatory system. Preferably, thepatch components resemble the viscoelastic properties of the skin andconform to the skin during movement to prevent undue shear anddelamination. Advantages of a patch comprising the topical formulationof the invention over conventional methods of administration include (i)that the dose is controlled by the patch's surface area, (ii) constantrate of administration, (iii) longer duration of action (the ability ofto adhere to the skin for 1, 3, 7 days or longer), (iv) improved patientcompliance, (v) non-invasive dosing, and (vi) reversible action (i.e.,the patch can simply be removed).

In certain embodiments, a patch suitable for use with the inventioncontains at least: (1) a backing layer and (2) a carrier formulated witha compound of the invention. Examples of patch systems suitable forpracticing the invention include, but are not limited to, matrix-typepatches; reservoir-type patches; multi-laminate drug-in-adhesive-typepatches; and monolithic drug-in-adhesive type-patch. See, for exampleGhosh, T. K.; Pfister, W. R.; Yum, S. I. Transdermal and Topical DrugDelivery Systems, Interpharm Press, Inc. p. 249-297, which isincorporated herein by reference in its entirety. These patches are wellknown in the art and generally available commercially.

The matrix patch comprises matrix containing an inventive compound, anadhesive backing film overlay, and preferably, but not necessarily, arelease liner. In some cases, it may be necessary to include aimpermeable layer to minimize drug migration into the backing film(e.g., U.S. Pat. No. 4,336,243, incorporated herein by reference). Incertain embodiments, the matrix containing the inventive compound isheld against the skin by the adhesive overlay. Examples of suitablematrix materials include but are not limited to lipophilic polymers,such as polyvinyl chloride, polydimethylsiloxane, and hydrophilicpolymers like polyvinylpyrrolidone, polyvinyl alcohol, hydrogels basedon gelatin, or polyvinylpyrrolidone/polyethylene oxide mixtures.Suitable release liners include but are not limited to occlusive,opaque, or clear polyester films with a thin coating of pressuresensitive release liner (e.g., silicone-fluorsilicone, andperfluorcarbon based polymers.

The reservoir type patch design is characterized by a backing filmcoated with an adhesive, and a reservoir compartment comprising a drugformulation preferably, in the form of a solution or suspension, that isseparated from the skin by a semipermeable membrane (e.g., U.S. Pat. No.4,615,699, incorporated herein by reference). The adhesive coatedbacking layer extends around the reservoir's boundaries to provide aconcentric seal with the skin and hold the reservoir adjacent to theskin.

The monolithic drug-in-adhesive patch design is characterized by theinclusion of the drug formulation in the skin contacting adhesive layer,a backing film and preferably, a release liner. The adhesive functionsboth to release the compound and adhere the compound matrix to the skin.The drug-in-adhesive system does not require an adhesive overlay andthus the patch size is minimized. Also, drug-in-adhesive type patchesare thin and comfortable (e.g., U.S. Pat. No. 4,751,087, incorporatedherein by reference).

The multi-laminate drug-in-adhesive patch design further incorporates anadditional semi-permeable membrane between two distinct drug-in-adhesivelayers or multiple drug-in-adhesive layers under a single backing film(Peterson, T. A. and Dreyer, S. J. Proceed. Intern. Symp. Control. Rel.Bioact. Mater. 21: 477-478, incorporated herein by reference).

Semi permeable membranes, useful with the reservoir or multi-laminatepatch, include thin non-porous ethylene vinyl acetate films or thinmacroporous films of polyethylene employed in microlaminate solid statereservoir patches.

Adhesives for use with the drug-in-adhesive type patches are well knownin the art and a practitioner skilled in the relevant art would know howto select an adhesive suitable for the intended use. Examples ofadhesives include, but are not limited to, polyisobutylenes, silicones,and acrylics. Preferably, adhesives can function under a wide range ofconditions, such as, high and low humidity, bathing, sweating etc.Preferably the adhesive is a composition based on natural or syntheticrubber; a polyacrylate such as, polybutylacrylate, polymethylacrylate,poly-2-ethylhexyl acrylate; polyvinylacetate; polydimethylsiloxane;pressure sensitive acrylic adhesives, for example Durotak.RTM. adhesives(e.g., Durotak.RTM. 2052, National Starch and Chemicals) or hydrogels(e.g., high molecular weight polyvinylpyrrolidone and oligomericpolyethylene oxide). The adhesive may contain a thickener, such as asilica thickener (e.g., Aerosil, Degussa, Ridgefield Park, N.J.) or acrosslinker such as, aluminumacetylacetonate.

Backing films may be occlusive or permeable and may be derived fromsynthetic polymers like polyolefin oils polyester, polyethylene,polyvinylidine chloride, and polyurethane or from natural materials likecotton, wool, etc. Occlusive backing films, such as syntheticpolyesters, result in hydration of the outer layers of the stratumcorneum while non-occlusive backings allow the area to breath (i.e.,promote water vapor transmission from the skin surface).

Selection of the appropriate dosage for the application site is animportant consideration. The rate of compound intradermal administrationfrom the topical formulation or patch is a function of skinpermeability, and skin permeability has been shown to vary betweenanatomical sites depending on the thickness of the stratum corneum. Forexample, the permeability, in general, increases in order from planterfoot arch, lateral ankle, palm, ventral forearm, dorsal forearm, back,chest, thigh, abdomen, scalp, axilla, forehead, and scrotum (Wester, R.C. and Maibach, H. I. (1989) Regional variation in PercutaneousAbsorption: In Percutaneous Absorption, Mechanism, Methodology, DrugDelivery, 2^(nd) ed., Eds. R. L. Bronaugh and H. I. Maibach, MarcelDekker, Inc., New York, pp. 111-119 (incorporated herein by reference)).Typically, the dosages and dosing frequency will be determined by atrained medical professional.

It will also be appreciated that the compounds and pharmaceuticalcompositions of the present invention can be formulated and employed incombination therapies, that is, the compounds and pharmaceuticalcompositions can be formulated with or administered concurrently with,prior to, or subsequent to, one or more other desired therapeutics ormedical procedures. The particular combination of therapies(therapeutics or procedures) to employ in a combination regimen willtake into account compatibility of the desired therapeutics and/orprocedures and the desired therapeutic effect to be achieved. It willalso be appreciated that the therapies employed may achieve a desiredeffect for the same disorder (for example, an inventive compound may beadministered concurrently with another immunomodulatory agent,anticancer agent or agent useful for the treatment of psoriasis), orthey may achieve different effects (e.g., control of any adverseeffects).

For example, other therapies or anticancer agents that may be used incombination with the inventive compounds of the present inventioninclude surgery, radiotherapy (in but a few examples, γ-radiation,neutron beam radiotherapy, electron beam radiotherapy, proton therapy,brachytherapy, and systemic radioactive isotopes, to name a few),endocrine therapy, biologic response modifiers (interferons,interleukins, and tumor necrosis factor (TNF) to name a few),hyperthermia and cryotherapy, agents to attenuate any adverse effects(e.g., antiemetics), and other approved chemotherapeutic drugs,including, but not limited to, alkylating drugs (mechlorethamine,chlorambucil, Cyclophosphamide, Melphalan, Ifosfamide), antimetabolites(Methotrexate), purine antagonists and pyrimidine antagonists(6-Mercaptopurine, 5-Fluorouracil, Cytarabile, Gemcitabine), spindlepoisons (Vinblastine, Vincristine, Vinorelbine, Paclitaxel),podophyllotoxins (Etoposide, Irinotecan, Topotecan), antibiotics(Doxorubicin, Bleomycin, Mitomycin), nitrosoureas (Carmustine,Lomustine), inorganic ions (Cisplatin, Carboplatin), enzymes(Asparaginase), and hormones (Tamoxifen, Leuprolide, Flutamide, andMegestrol), to name a few. For a more comprehensive discussion ofupdated cancer therapies see, The Merck Manual, Seventeenth Ed. 1999,the entire contents of which are hereby incorporated by reference. Seealso the National Cancer Institute (CM) website (www.nci.nih.gov) andthe Food and Drug Administration (FDA) website for a list of the FDAapproved oncology drugs (www.fda.gov/cder/cancer/druglistframe—SeeAppendix A).

In certain embodiments, the pharmaceutical compositions of the presentinvention further comprise one or more additional therapeutically activeingredients (e.g., chemotherapeutic and/or palliative). For purposes ofthe invention, the term “Palliative” refers to treatment that is focusedon the relief of symptoms of a disease and/or side effects of atherapeutic regimen, but is not curative. For example, palliativetreatment encompasses painkillers, antinausea medications andanti-sickness drugs. In addition, chemotherapy, radiotherapy and surgerycan all be used palliatively (that is, to reduce symptoms without goingfor cure; e.g., for shrinking tumors and reducing pressure, bleeding,pain and other symptoms of cancer).

In certain embodiments, compounds of the invention are useful for thetreatment of psoriasis and pharmaceutical compositions containing themmay be administered in combination with any of the antipsoriatictherapies or therapeutic agents known in the art. For example, therapiesor antipsoriatic agents that may be used in combination with theinventive compounds of the present invention include Ultraviolet lighttreatment (e.g., sunlight), lubricants, keratolytics, emollients (e.g.,Aqueous Cream, E45, and Emulsifying ointment), ammoniated mercury,topical vitamin D analogs (e.g., Calcipotriol (Dovonex), Tacalcitol(Curatoderm)), dithranol (e.g., Dithrocream and Miconal), tar (e.g.,Alphosyl, anthralin), topical steroids (e.g., corticosteroids,halobetasol), topical retinoids (e.g., zorac, Tazarotene), systemicantimetabolites (e.g., oral methotrexate), immunosuppressive drugs(e.g., oral cyclosporine, tacrolimus, mycophenolate, and mofetil) andoral retinoids (e.g., acitretin).

Treatment Kits

In other embodiments, the present invention relates to a kit forconveniently and effectively carrying out the methods in accordance withthe present invention. In general, the pharmaceutical pack or kitcomprises one or more containers filled with one or more of theingredients of the pharmaceutical compositions of the invention. Suchkits are especially suited for the delivery of solid oral forms such astablets or capsules. Such a kit preferably includes a number of unitdosages, and may also include a card having the dosages oriented in theorder of their intended use. If desired, a memory aid can be provided,for example in the form of numbers, letters, or other markings or with acalendar insert, designating the days in the treatment schedule in whichthe dosages can be administered. Alternatively, placebo dosages, orcalcium dietary supplements, either in a form similar to or distinctfrom the dosages of the pharmaceutical compositions, can be included toprovide a kit in which a dosage is taken every day. Optionallyassociated with such container(s) can be a notice in the form prescribedby a governmental agency regulating the manufacture, use or sale ofpharmaceutical products, which notice reflects approval by the agency ofmanufacture, use or sale for human administration.

EQUIVALENTS

The representative examples that follow are intended to help illustratethe invention, and are not intended to, nor should they be construed to,limit the scope of the invention. Indeed, various modifications of theinvention and many further embodiments thereof, in addition to thoseshown and described herein, will become apparent to those skilled in theart from the full contents of this document, including the exampleswhich follow and the references to the scientific and patent literaturecited herein. It should further be appreciated that the contents ofthose cited references are incorporated herein by reference to helpillustrate the state of the art.

The following examples contain important additional information,exemplification and guidance that can be adapted to the practice of thisinvention in its various embodiments and the equivalents thereof.

EXEMPLIFICATION

The compounds of this invention and their preparation can be understoodfurther by the examples that illustrate some of the processes by whichthese compounds are prepared or used. It will be appreciated, however,that these examples do not limit the invention. Variations of theinvention, now known or further developed, are considered to fall withinthe scope of the present invention as described herein and ashereinafter claimed.

1) General Description of Synthetic Methods:

The practitioner has a a well-established literature of macrolidechemistry to draw upon, in combination with the information containedherein, for guidance on synthetic strategies, protecting groups, andother materials and methods useful for the synthesis of the compounds ofthis invention.

The various references cited herein provide helpful backgroundinformation on preparing compounds similar to the inventive compoundsdescribed herein or relevant intermediates, as well as information onformulation, uses, and administration of such compounds which may be ofinterest.

Moreover, the practitioner is directed to the specific guidance andexamples provided in this document relating to various exemplarycompounds and intermediates thereof.

The compounds of this invention and their preparation can be understoodfurther by the examples that illustrate some of the processes by whichthese compounds are prepared or used. It will be appreciated, however,that these examples do not limit the invention. Variations of theinvention, now known or further developed, are considered to fall withinthe scope of the present invention as described herein and ashereinafter claimed.

According to the present invention, any available techniques can be usedto make or prepare the inventive compounds or compositions includingthem. For example, a variety of solution phase synthetic methods such asthose discussed in detail below may be used. Alternatively oradditionally, the inventive compounds may be prepared using any of avariety combinatorial techniques, parallel synthesis and/or solid phasesynthetic methods known in the art.

It will be appreciated as described below, that a variety of inventivecompounds can be synthesized according to the methods described herein.The starting materials and reagents used in preparing these compoundsare either available from commercial suppliers such as Aldrich ChemicalCompany (Milwaukee, Wis.), Bachem (Torrance, Calif.), Sigma (St. Louis,Mo.), or are prepared by methods well known to a person of ordinaryskill in the art following procedures described in such references asFieser and Fieser 1991, “Reagents for Organic Synthesis”, vols 1-17,John Wiley and Sons, New York, N.Y., 1991; Rodd 1989 “Chemistry ofCarbon Compounds”, vols. 1-5 and supps, Elsevier Science Publishers,1989; “Organic Reactions”, vols 1-40, John Wiley and Sons, New York,N.Y., 1991; March 2001, “Advanced Organic Chemistry”, 5th ed. John Wileyand Sons, New York, N.Y.; and Larock 1990, “Comprehensive OrganicTransformations: A Guide to Functional Group Preparations”, 2^(nd) ed.VCH Publishers. These schemes are merely illustrative of some methods bywhich the compounds of this invention can be synthesized, and variousmodifications to these schemes can be made and will be suggested to aperson of ordinary skill in the art having regard to this disclosure.

The starting materials, intermediates, and compounds of this inventionmay be isolated and purified using conventional techniques, includingfiltration, distillation, crystallization, chromatography, and the like.They may be characterized using conventional methods, including physicalconstants and spectral data.

Certain exemplary compounds of the invention are listed below and arereferred to by compound number as indicated.

General Reaction Procedures:

Unless mentioned specifically, reaction mixtures were stirred using amagnetically driven stirrer bar. An inert atmosphere refers to eitherdry argon or dry nitrogen. Reactions were monitored either by thin layerchromatography, by proton nuclear magnetic resonance (NMR) or byhigh-pressure liquid chromatography (HPLC), of a suitably worked upsample of the reaction mixture.

Listed below are abbreviations used for some common organic reagentsreferred to herein:

m-CPBA: meta-chloroperbenzoic acid

DDQ: 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone

DEAD Diethyl azodicarboxylate

DIBAL-H: Diisobutyl aluminum hydride

DMAP: N,N-Dimethylaminopyridine

DMF: N,N-Dimethylformamide

HMPA: Hexamethylphosphoramide

LDA: Lithium diisopropyl amide

LiHMDS: Lithium bis(trimethylsilyl)amide

PCC: Pyridinium chlorochromate

TBAF: Tetrabutylammonium fluoride

THF: Tetrahydrofuran

General Work Up Procedures:

Unless mentioned specifically, reaction mixtures were cooled to roomtemperature or below then quenched, when necessary, with either water ora saturated aqueous solution of ammonium chloride. Desired products wereextracted by partitioning between water and a suitable water-immisciblesolvent (e.g. ethyl acetate, dichloromethane, diethyl ether). Thedesired product containing extracts were washed appropriately with waterfollowed by a saturated solution of brine. On occasions where theproduct containing extract was deemed to contain residual oxidants, theextract was washed with a 10% solution of sodium sulphite in saturatedaqueous sodium bicarbonate solution, prior to the aforementioned washingprocedure. On occasions where the product containing extract was deemedto contain residual acids, the extract was washed with saturated aqueoussodium bicarbonate solution, prior to the aforementioned washingprocedure (except in those cases where the desired product itself hadacidic character). On occasions where the product containing extract wasdeemed to contain residual bases, the extract was washed with 10%aqueous citric acid solution, prior to the aforementioned washingprocedure (except in those cases where the desired product itself hadbasic character). Post washing, the desired product containing extractswere dried over anhydrous magnesium sulphate, and then filtered. Thecrude products were then isolated by removal of solvent(s) by rotaryevaporation under reduced pressure, at an appropriate temperature(generally less than 45° C.).

On occasions where triphenylphosphine oxide was a major byproduct of thereaction, the reaction mixture was added directly to a large volume ofwell-stirred hexane. The resultant precipitate of triphenylphosphineoxide was removed by filtration and the filtrate processed in the usualmanner.

General Purification Procedures:

Unless mentioned specifically, chromatographic purification refers toflash column chromatography on silica, using a single solvent or mixedsolvent as eluent. Suitably purified desired product containing eluteswere combined and concentrated under reduced pressure at an appropriatetemperature (generally less than 45° C.) to constant mass. Finalcompounds were dissolved in 50% aqueous acetonitrile, filtered andtransferred to vials, then freeze-dried under high vacuum beforesubmission for biological testing.

Synthesis for Commonly Used Intermediates:

Starting material (50.0 g, 0.27 mol) was dissolved in 650 mL of THF at0° C. Triphenylphosphine (93.6 g, 0.35 mol) was added, followed bymethanol (12.2 mL, 0.30 mol) and diethyl azodicarboxylate (56.2 ml, 0.35mol). After stirring at 0° C. for 1.5 h, the reaction mixture wasconcentrated, redissolved in diethyl ether, washed with 1N sodiumhydroxide solution. The aqueous layer was acidified with concentratedhydrochloric acid and extracted with diethyl ether. After purificationon silica gel column, 42.0 g of 509-HD-207 was obtained as a pale yellowsolid in 78% yield.

To the reaction flask containing NaH (95%, 14.5 g, 0.57 mol) in 1 L ofTHF at 0° C. was added 509-HD-207 (75.0 g, 0.38 mol) in 0.5 L of THF.After stirred for 0.5 h, chloromethyl methyl ether (43.6 mL, 0.57 mol)was added. After stirred at 0° C. for 1 h, it was warmed up to roomtemperature. The reaction was quenched with water and extracted withpentane. After purification on silica gel column, 83 g of 509-HD-209 wasobtained as colorless oil in 92% yield.

Diisopropyl amine (68.1 mL, 486 mol) was dissolved in 1 L of THF at 0°C. n-BuLi (2.5 M, 207 mL) was added. The solution was stirred for 20min, and then cooled down to −78° C. The solution of 509-HD-209 (77.8 g,324 mol) in 250 mL of THF was added slowly. 1 h later the solution ofdiphenyl diselenide (85.9 g, 275 mol) in 250 ml of THF was added. Afterstirring at −78° C. for 1 h, the reaction was quenched with saturatedammonium chloride solution, and extracted with diethyl ether. Afterpurification on silica gel column, 90.2 g of 509-HD-211 was obtained aspale yellow oil in 68% yield.

509-HD-211 (90.2 g, 228 mmol) was dissolved in 500 mL of ethanol. Sodiumhydroxide solution (1N, 456 mL) was added. The resulting solution washeated under reflux for 12 h. The reaction mixture was acidified with 1Nhydrochloric acid, extracted with diethyl ether and concentrated, giving84.6 g of 509-HD-212 as a pale yellow solid in 97% yield.

509-HD-212 (84.6 g, 222 mmol) and triphenylphosphine (75.7 g, 289 mmol)was dissolved in a mixture of 500 mL of diethyl ether and 125 mL oftoluene at 0° C. 2-(trimethylsilyl)ethanol (38.2 mL, 266 mmol) anddiethyl azodicarboxylate (45.4 mL, 289 mmol) were added respectively.After stirred for 10 min, it was warmed up to room temperature. Largeamount of pentane was added to precipitate the solid. After filtration,the crude product was purified on silica gel column and 80.0 g of509-HD-213 was obtained as pale yellow oil in 75% yield.

To a solution of starting material (157 g, 0.86 mol) in 1.6 L of toluene(slightly cloudy), TMS-ethanol (150 g, 1.27 mol, 1.48 eq.), and PPh₃(440 g, 1.68 mol, 1.95 eq.) were added. After cooled to 0° C., DEAD (725mL of 40% solution, 1.29 mol, 1.5 eq.) was slowly added by droppingfunnel while maintaining internal temp below 10° C. in three hours.After stirred at rt for 48 h, it was poured into a rapid stirredsolution of hexanes (12 L). The solid was filtered through a celite padand the pad was washed with 1 L of hexanes. The filtrates were combinedand concentrated to give the crude product as brown oil. The oil waspurified on silica gel with hexanes/EtOAc (15:1, 10:1, 6:1) to give 140g of product as an off white solid.

The phenol (140 g, 0.49 mol) was dissolved in 400 mL of CH₂Cl₂, DBU(135.5 mL, 0.91 mol, 1.85 eq.) was added. After cooled to 0° C., MOMCl(66 mL, 0.87 mol, 1.78 eq.) was added. After stirred at rt for 24 h, itwas quenched with Sat. NH₄Cl, extracted with EtOAc. The organic layerwas washed with brine, dried and concentrated to dryness. The crudeproduct was purified on column with hexanes/EtOAc, 10:1, 6:1 to give 132g of desired product (82% in yield).

To a solution of diisopropylamine (143.6 mL, 1.02 mol, 2.3 eq.) in 320mL of THF, n-BuLi (422.6 mL, 2.5 M) was added at 0° C. by additionfunnel, while controlling the internal temperature around 5° C. After 5min at that temperature, the reaction was cooled to −78° C. A solutionof starting material (145 g, 0.44 mol) in 475 mL of THF was added byaddition funnel while internal temperature controlled at or below −70°C. After addition, it was stirred at −70° C. for 30 min. Then at thattemperature, a solution of PhSe₂ (140 g, 0.45 mol, 1 eq.) in 400 mL ofTHF was added by addition funnel. After addition, it was stirred at −78°C. for 45 min. The reaction was quenched with sat. NH₄Cl, diluted withEtOAc. It was warmed to rt. The organic layer was washed with brine,dried and concentrated to dryness. The crude product was used withoutpurification for next step.

The crude product from last step was dissolved in 300 mL of EtOH. Then300 mL of 1N NaOH was added. The reaction was heated at 80° C.overnight. After cooled, it was transferred to a separatory funnel andwashed with hexanes. The aq. Layer was acidified at 0° C. with 1N HCl topH=3. Then it was extracted with EtOAc (3×). The combined organic layerswere washed with brine, dried and concentrated to dryness. The crudeacid was used for next step without purification.

The esterification was run according to the first step using DEAD (200g), PPh₃ (330 g) and TMS-ethanol (150 g) in 1.4 L of toluene to give 145g of product after column chromatography.

Other C14-substitutions aromatic pieces such as methyl, ethyl, Benzyl,PMB (MPM) etc. were made in analogues manner by substituting the firststep with corresponding alcohols such as methanol, ethanol, benzylalcohol, or PMB alcohol etc.

To a stirring solution of diisopropylamine (2 eq., 366 mmol, 51.3 mL) indry THF (200 mL) at −78° C. was added slowly a 1.6 M solution of n-BuLi(2 eq., 366 mmol, 230 mL) over a period of 20 min. The reaction mixturewas warmed to 0° C. and allowed to stir for 45 min after which thesolution was cooled back to −78° C. Then, a solution ofmethyl-3-hydroxybutyrate (21.6 g, 183 mmol) in thy THF (100 mL) wasadded slowly over a period of 20 min after which neat MeI (5 eq, 915mmol, 57 mL) was added over a period of 5 min. The reaction mixture wasallowed to stir for 10 min at −78° C. then warmed to rt, and stirred for2 h. The reaction was quenched with a saturated solution of NH₄Cl (350mL), extracted with Et₂O (3×400 mL), the combined organic extracts werewashed with a saturated solution of NH₄Cl (350 mL), water (2×450 mL),brine (450 mL), dried with K₂CO₃, filtered and concentrated. The crudealcohol 555-RB-224 was dissolved in dry DMF (100 mL), imidazole (2.5 eq,363 mmol, 24.7 g) was added and the mixture was cooled to 0° C. inice/water bath. Then TBSCl (1.2 eq, 33.0 mmol, 5 g) was added, themixture was allowed to warm slowly to rt and stirred for 16 h afterwhich a saturated solution of NaHCO₃ (250 mL) was added. The mixture wasextracted with Et₂O (3×250 mL) and the combined organic extracts werewashed with a saturated solution of NaHCO₃ (350 mL), water (3×350 mL),brine (350 mL), dried with Na₂SO₄, filtered and concentrated. The crudeproduct was purified by chromatography on silica gel using 5%EtOAc/hexane to give 31.7 g (129 mmol, 70% 2 steps) of the protectedcompound 554-RB-225.

In a 5 L three neck flask equipped with mechanical stirring was placed554-RB-225 (221 mmol, 54.5 g) dissolved in toluene (750 mL). The mixturewas cooled to −78° C. and DIBAL-H (1M in toluene, 2.5 eq., 553 mmol, 553mL) was added slowly. The mixture was allowed to stir at −78° C. for 15min after which it was warmed to 0° C. and stirred for 2 h. Reaction wasquenched with MeOH (10 eq., 2.2 mol, 89 mL) at −78° C. and allowed towarm to rt, Et₂O (2.5 L) and a saturated solution of Na₂SO₄ (1 L) wereadded and the solution was stirred overnight. Mixture was filteredthrough celite and the solid was washed with Et₂O (2×1 L). The filtratewas concentrated under reduced pressure and the resulting residue waspurified by chromatography on silica gel using 10-15% EtOAc/hexane togive 40.6 g (186 mmol, 76%) of alcohol 554-RB-227.

To a solution of oxalyl chloride (2 eq., 372 mmol, 33.0 mL) in CH₂Cl₂(800 mL), DMSO (4 eq., 744 mmol, 53.0 mL) was added at −78° C. After 30min at −78° C., a solution of alcohol 554-RB-227 (186 mmol, 40.6 g) inCH₂Cl₂ (200 mL) was added over a period of 15 min. After 50 min at −78°C., Et₃N (4 eq., 744 mmol, 104 mL) was added and the reaction was warmedto 0° C. and stirred for 45 min. It was quenched with a saturatedsolution of NH₄Cl (500 mL), extracted with EtOAc (1×2 L, 2×400 mL). Thecombined organic layers were dried with Na₂SO₄, filtered andconcentrated. The crude aldehyde was filtered through a short silica gelcolumn with 10% EtOAc/hexane. To a solution of PPh₃ (3 eq., 558 mmol,146 g) in CH₂Cl₂ (2 L), CBr₄ (1.5 eq., 279 mmol, 92.5 g) was added at 0°C. Then, a solution of aldehyde and Et₃N (1 eq., 186 mmol, 26 mL) inCH₂Cl₂ (200 mL) was added. The solution was stirred under nitrogen at rtfor one hour after which the mixture was concentrated under reducedpressure. The residue was dissolved in CH₂Cl₂, poured into hexane (2.5L) and stirred. The precipitate was filtered through celite and thefiltrate was concentrated. Purification by chromatography on silica gelusing CH₂Cl₂ as eluent gave 62.9 g (169 mmol, 91% 2 steps) of554-RB-228.

A 5 L, 3-neck flask was equipped with mechanical stirring, cooling bathand flushed with nitrogen. Then, a solution of alcohol 491-HAD-46 (202mmol, 52.5 g) and MPMOTCl (2 eq., 404 mmol, 115.5 g) in Et₂O (1 L) wasadded to the flask and cooled to 0° C. A solution of TfOH (1.5 mL) inEt₂O (120 mL) was added slowly with a syringe pump over a period of 50min. Then, a saturated solution of NaHCO₃ (500 mL) was added, themixture was extracted with Et₂O (2×700 mL); the combined organicextracts were washed with brine (2×1 L), dried over Na₂SO₄ andconcentrated. The residue was dissolved in CH₂Cl₂, poured in hexane (2.5L), the precipitated was filtered through celite and the filtrate wasconcentrated under reduced pressure. The crude material was purified bychromatography on silica gel using 5-10% EtOAc/hexane as eluent to give57.3 g (151 mmol, 75%) of the protected alcohol 554-RB-235.

In a 3 L, three neck flask equipped with mechanical stirring was placed554-RB-235 (151 mmol, 57.3 g) dissolved in toluene (750 mL). The mixturewas cooled to −78° C. and DIBAL-H (1M in toluene, 2.65 eq., 400 mmol,400 mL) was added slowly. The mixture was allowed to stir at −78° C. for10 min after which it was warmed to 0° C. and stirred for 20 min.Reaction was quenched with MeOH (10 eq., 1.5 mol, 61 mL) at −78° C. andallowed to warm to rt. Et₂O (2.5 L) and a saturated solution of Na₂SO₄(1 L) were added and the solution was stirred overnight. Mixture wasfiltered through celite and the solid was washed with Et₂O (2×1 L). Thefiltrate was concentrated under reduced pressure and the resultingresidue was purified by chromatography on silica gel using 20-40%EtOAc/hexane to give 27 g (91 mmol, 60%) of alcohol 554-RB-237.

To a solution of oxalyl chloride (2 eq., 202 mmol, 18 mL) in CH₂Cl₂ (650mL), DMSO (4 eq., 404 mmol, 29 mL) was added at −78° C. After 30 min at−78° C., a solution of alcohol 554-RB-237 (101 mmol, 30 g) in CH₂Cl₂(100 mL) was added over a period of 30 min. After 45 min at −78° C.,Et₃N (4 eq., 404 mmol, 56 mL) was added and the reaction was warmed to0° C. and stirred for 45 min. It was quenched with a saturated solutionof NH₄Cl (250 mL), extracted with EtOAc (1×2 L, 2×250 mL). The combinedorganic layers were dried with Na₂SO₄, filtered and concentrated. Thecrude aldehyde was purified by chromatography on silica gel with 10%EtOAc/hexane to give 27 g (91.7 mmol, 91%) of aldehyde 554-RB-238.

Dibromoolefin 554-RB-228 (1.5 eq., 138 mmol, 51.2 g) was dissolved inTHF (1 L) and cooled to −78° C., under nitrogen. Then, n-BuLi(1.6M/hexane, 3.3 eq., 302 mmol, 189 mL) was added and the reaction wasstirred at −78° C. for 40 min, at 0° C. for 30 min, then cooled back to−78° C. Aldehyde 554-RB-238 (91.7 mmol, 27.0 g) dissolved in THF (200mL) was added to the solution and stirred for 30 min at −78° C. Thesolution was allowed to warm to rt and was stirred for 1.5 hrs. Themixture was quenched with water (700 mL), extracted with EtOAc (3×750mL) and the combined organic extracts were washed with brine (1 L),dried with Na₂SO₄, filtered and concentrated. The residue was purifiedby chromatography on silica gel using 10-30% EtOAc/hexane to give 43.7 g(86 mmol, 94%) of 554-RB-240.

554-RB-240 (86 mmol, 43.7 g) was dissolved in hexane (1 L). Then,quinoline (1 mL) and Lindlar catalyst (10 g) were added. H₂ balloon wasmounted and the mixture was purged 5× with H₂. Reaction was stirredunder hydrogen. After 13 hrs, 17 hrs, 22 hrs, and 38 hrs, catalyst wasfiltered and new catalyst (10 g) and quinoline (1 mL) were added eachtime. After 42 hrs, reaction was stopped, catalyst was filtered throughcelite and mixture was concentrated under reduced pressure. Then, crude554-RB-241 was dissolved in CH₂Cl₂ (700 mL), Et₃N (3.75 eq., 323 mmol,45 mL), BzCl (3 eq., 258 mmol, 30 mL) and DMAP (0.075 eq., 6.45 mmol,788 mg) were added and the mixture was stirred for 96 hrs at rt undernitrogen. The mixture was diluted with EtOAc (2 L) and a 0.1N solutionof NaOH (800 mL). Organic layer was separated and the aqueous phase wasextracted with EtOAc (2×500 mL). The organic combined extracts werewashed with a 0.2N solution of NaOH (5×500 mL), brine, dried withNa₂SO₄, filtered and concentrated. The crude compound was filteredthrough a silica gel column with 5% EtOAc/hexane to give a quantitativeyield of protected compound 554-RB-242.

554-RB-242 was dissolved in CH₂Cl₂ (500 mL), H₂O (250 mL) and DDQ (1.1eq., 94.6 mmol, 21.3 g) were added and the mixture was stirredvigorously at rt for 4 hrs. The mixture was quenched with a 0.2Nsolution of NaOH (500 mL) and diluted with EtOAc (2 L). The organiclayer was separated and the aqueous phase was back extracted with EtOAc(2×500 mL). The combined organic layers were washed with a 0.2N solutionof NaOH (3×700 mL), brine (700 mL), dried with Na₂SO₄, filtered andconcentrated. The crude residue was purified by chromatography on silicagel using 10% EtOAc/hexane to give 39.9 g (81 mmol, 94% 3 steps fromacetylene) of free alcohol 554-RB-244.

To a solution of 554-RB-244 (6.09 mmol, 3.0 g) in toluene (100 mL), Ph₃P(1.7 eq., 10.4 mmol, 2.71 g) was added at rt. Then, CH₃I (1.3 eq., 7.92mmol, 0.49 mL) and DEAD (1.1 eq., 6.70 mmol, 1.45 mL) were added at thesame time. The mixture was stirred for 1.5 hrs at rt after which it waspoured in hexane and stirred for 10 min. The precipitate was filteredand the filtrate was concentrated. The residue was purified bychromatography on silica gel using 5% EtOAc/hexane to give 3.40 g (5.64mmol, 93%) of iodide 554-RB-260.

Preparation of C4-H Series Acyclic Segment:

To a solution of 531-YW-2-3 (7.5 g) in 20 mL of DMF, imidazole (5 g) andTBDPSCl (8.4 g) were added. During the addition exotherm was observed.After 3 h, it was diluted with EtOAc, washed with aq. Sat. NH₄Cl andbrine. After drying and filtration, it was concentrated. The crudeproduct was purified on silica gel with Hexanes/EtOAc, 20:1, and 10:1 togive 11.0 g of desired product.

The product was dissolved in 200 mL of THF. LAH (1.4 g) was added at 0°C. After 10 min at 0° C., it was quenched with water, 1N NaOH. Themixture was stirred at rt for 1 h. Then it was filtered. The combinedfiltrates were concentrated to dryness. The crude product was purifiedon silica gel with Hexanes/EtOAc, 10:1, 4:1, and 2:1 to give 9.0 g ofdesired product with satisfactory 111 NMR spectra, 343-YW-275.

To a solution of commercial available (s)-3-hydroxy butanol (10 g,Aldrich) in 50 mL of DMF, TsOH (20 mg, catalytic) and MeOPhCH(OMe)₂ (24g) were added. After 3 h at 35° C. on a rotovap with slight vacuum, itwas cooled and quenched with aq. Sat. NaHCO₃. The mixture was extractedwith EtOAc (3×). The organic layers were washed with brine (2×), driedand concentrated. The crude product was evaporated with toluene (3×).

The crude product was dissolved in 700 mL of CH₂Cl₂. At 0° C., DIBAL-Hsolution (200 mL, 1.0 M, excess) was added. The reaction was warmed toroom temperature overnight. Then it was quenched with methanol (50 mL),sat. Na₂SO₄ at 0° C. The mixture was diluted with Et₂O (1.5 L). Afterstirred for 5 h, it was filtered through a pad of celite. The filtratewas concentrated to give an oil. The oil was purified on silica gel withHexanes/EtOAc, 10:1, 6:1, 3:1, and 1:1 to give 24 g of desired product,343-YW-203.

To a solution of DMSO (3.7 mL) in 150 mL of CH₂Cl₂, a solution of343-W-203 (3.6 g) in 50 mL of CH₂Cl₂ was added. At 0° C., solid P₂O₅(6.06 g) was added. The reaction was warmed to room temperature. Afterstirred at room temperature for 1 h, the reaction turned light pink andwas cooled to 0° C. Et₃N (12 mL) was added. After 15 min at 0° C., itwas warmed to room temperature. After 10 min, it was quenched with sat.NH₄Cl, and extracted with CH₂Cl₂ (2×). The organic layers were dried andconcentrated. The crude product was suspended in Et₂O and filtered. Thefiltrates were concentrated to dryness.

To a solution of PPh₃ (11.6 g) in 30 mL of CH₂Cl₂, CBr₄ (7.4 g) wasadded at 0° C. The internal temperature was controlled below 10° C.After 10 min, a solution of aldehyde in 20 mL of CH₂Cl₂ was added. Theinternal temperature went up to 20° C. It was warmed to room temperatureand stirred for 1 h. Then it was poured into a rapid stirring pentanesolution. The precipitation was filtered. The filtrates wereconcentrated. The crude product was purified on silica gel column withhexanes/EtOAc, 20:1, 15:1, 10:1 to give 4.4 g of the desired product,343-W-276.

Oxidation of Alcohol:

To a solution of 343-YW-275 (3.6 g) in 25 mL of CH₂Cl₂, DMSO (1.84 mL)was added followed by P₂O₅ (3.65 g) solid at 0° C. Then it was warmed toroom temperature for 1 h. After cooled to 0° C., Et₃N (5.94 mL) wasadded. After stirred at 0° C. for 30 min, it was warmed to roomtemperature. After stirred at room temperature for 3 h, it was quenchedwith sat. NH₄Cl, and extracted with CH₂Cl₂ (2×). The organic layers werewashed with brine, dried and concentrated. The crude product waspurified on silica gel with Hexanes/EtOAc, 10:1, 6:1, and 4:1 to give2.5 g of aldehyde, 343-YW-277.

Coupling:

To a solution of 343-YW-276 (4.5 g, 12.36 mmol, 2 eq.) in 35 mL of THF,n-BuLi (5.4 mL, 2.5M, 2.2 eq.) was added at −78° C. After 10 min at −78°C., it was warmed to room temperature for 30 min. After cooled back to−78° C., a solution of 343-YW-277 (2.5 g, 6.06 mmol, 1 eq.) in 10 mL ofTHF was added. It was then warmed to 0° C., after 30 min. After 4 h at0° C., it was quenched with aq. Sat. NH₄Cl, extracted with EtOAc (2×).The organic layers were washed with brine, dried and concentrated. Thecrude product was purified on silica gel with Hexanes/EtOAc, 20:1, 15:1,10:1, 6:1 to give 2.5 g of desired product, 343-YW-278 along withrecovered 343-YW-276.

To a solution of 343-YW-278 (2.50 g) in 100 mL of hexanes, Lindlarcatalyst (330 mg, catalytic) and quinoline (50 μL, catalytic) wereadded. After degassed under vacuum and refilled with H₂ for severaltimes, it was stirred under hydrogen balloon for 3 h. Then the catalystwas filtered and fresh catalyst was added. After degassing, it wasstirred under hydrogen overnight. The reaction was filtered throughcelite. The filtrates were combined and concentrated to dryness to give2.4 g of desired product, 343-YW-279.

To a solution of 343-YW-279 (2.4 g) in 15 mL of CH₂Cl₂, BzCl (0.9 mL),Et₃N (2 mL) and DMAP (50 mg) were added. After 18 h, another 200 uL ofBzCl was added. After total 24 h, it was quenched with aq. Sat. NH₄Cland extracted with EtOAc (2×). The organic layers were washed withbrine, dried and concentrated. The crude product was purified on silicagel with Hexanes/EtOAc, 20:1, 10:1, and 6:1 to give 2.2 g of desiredester.

To a solution of the ester from last step in 10 mL of THF, solid TBAFwas added. After 18 h, it was quenched with sat. NH₄Cl and extractedwith EtOAc. The organic layer was washed with brine, dried andconcentrated. The crude product was purified on silica gel withHexanes/EtOAc, 10:1, 6:1, 4:1, and 2:1 to give 1.45 g of alcohol.

To a solution of the alcohol from last step and PPh₃ (1.3 g) in 20 mL oftoluene, DEAD (750 μL) and MeI (250 μL) were added simultaneously atroom temperature. After 30 min, it was diluted with CH₂Cl₂ to a clearsolution. Then it was poured into pentanes with rapid stirring. Theprecipitation was filtered through a celite pad. The filtrate wasconcentrated. The crude product was purified on silica gel withHexanes/EtOAc, 20:1, 10:1, 8:1 to give 1.5 g of desired product,343-YW-281. A satisfactory ¹H NMR was obtained.

Preparation of Advance Phenol Intermediate for C14-Analog Synthesis:

Iodide 2 (4.94 g, 8.2 mmol, 1.0 eq.) and selenide 3 (7.00 g, 12.3 mmol,1.5 eq.) were dissolved in HMPA (10.0 mL) and THF (90 mL). The resultingmixture was magnetically stirred at −78′C and slowly added over 50 minLiHMDS (18.0 mL, 9.0 mmol, 1.1 eq., LiHMDS 0.5M in THF, addition rate of0.32 mL/min). The reaction mixture was stirred 1 h and 45 min at −78°C., quenched with NH₄Cl sat. (200 mL), diluted with H₂O, added ethylacetate (500 mL). The layers were separated and the aqueous one wasextracted with ethyl acetate (2×500 mL). The combined organic layerswere washed with H₂O (2×300 mL), dried with Na₂SO₄, filtered, andconcentrated under reduced pressure. The crude oil was purified on aSiO₂ column (230-400 Mesh silica). The products were dissolved in hexaneprior to be loaded on the column. Elution: 3%, then 5% ethylacetate/hexane. The desired product 4 (5.43 g, 64% yield) was isolatedas viscous brownish oil.

The crude substrate 4 (mixture of selenide 3 and coupled material 4,<58.9 mmol) was dissolved in CH₂Cl₂ (750 mL), cooled down to 0° C. andadded in small portions MCPBA (Aldrich 57-86%, 43.5 g, >144 mmol, 2.4eq.).

The first portions are exothermic and towards the end of the addition,no exotherm was noticed, Tmax was 4° C. Stirred for 45 min at 0° C.,triethylamine (50 mL, 357 mmol, 6 eq.) was added SLOWLY! EXOTHERMIC!Tmax=10° C. Once the exotherm had ceased, the reaction mixture waswarmed up to it. Stirred for 60 min at that temperature, a solution ofNaS₂O₃ (51.5 g) was prepared using NaHCO₃ saturated and distilled water.Layers were separated; the aqueous one was extracted twice with CH₂Cl₂.The combined organic layers were dried with Na₂SO₄, filtered,concentrated down, added crude material from a previous small-scale run,purified on a SiO₂ column (1.25 kg of 230-400 Mesh silica fromSilicycle). The crude material was loaded on the column as a slurryprepared with 3% ethyl acetate/hexane and 230-400 Mesh silica. Elution:3% (8 L), 7.5% (8 L) and 10% (6 L) ethyl acetate/hexane. The desiredmaterial 5 (16.3 g, 30% combined yield since the selenide coupling) isviscous oil.

Substrate 5 was dissolved in THF (43 mL), added an imidazole.HClbuffered TBAF solution (60.5 mL, 60.5 mmol of TBAF, 3.4 eq. of TBAF 1 Min THF and 45 mmol of imidazole.HCl, 2.5 eq. of imidazole.HCl). Thatbuffered solution was prepared as follows: imidazole.HCl was dissolvedin a commercial 1 M TBAF/THF solution to give a resulting imidazole.HClmolarity of 0.75 M. The resulting reaction mixture was stirred 2 min atrt then it was added drop-wisely a regular TBAF solution (76 mL, 76mmol, TBAF 1.0 M in THF). The reaction mixture was stirred in an oilbath at 50° C. during a total of 88 h, cooled down to rt, added NH₄Clsat. (300 mL) and Et₂O (300 mL). The layers were separated and theaqueous was extracted with Et₂O (3×150 mL). The organic layers werecombined, washed with brine (3×100 mL), dried with Na₂SO₄, filtered,concentrated to dryness, azeotroped twice with Et₂O (2×100 mL) givingdesired 6 that was used crude for the next step.

In a three neck 5 L flask equipped with a condenser and an additionfunnel, was added CH₂Cl₂ (2 L) followed by triethylamine (7.6 mL, 54.0mmol, 3.0 eq.) and 2-chloro-1-methylpyridinium iodide (14.2 g, 54.0mmol, 3.0 eq.). The resulting mixture was warmed up to reflux and addeddrop-wisely a CH₂Cl₂ solution of hydroxy-acid 6 (14.4 g of crudematerial in 230 mL of CH₂Cl₂). The addition took 3 h and the resultingreaction mixture was stirred at reflux for 12 h, then cooled down to it,salts were filtered and CH₂Cl₂ was removed under reduced pressure. Theresidue was dissolved in Et₂O, washed with a 1:1 mixture of saturatedbrine and saturated NaHCO₃. The aqueous layer was extracted twice withEt₂O. The combined organic layers were washed with brine, dried withNa₂SO₄ and concentrated under reduced pressure, purified on a SiO₂column (250 g of 230-400 Mesh silica from Silicycle). The crude materialwas dissolved in CH₂Cl₂ prior to be loaded on the column. Elution: 15%,25%, 35% ethyl acetate/hexane. The desired macrocycle 7 (7 g) was stillslightly contaminated was used directly for the next step.

To a solution of 7 (7 g, <18 mmol, 1.0 eq.) in THF (40 mL) at rt, wasadded drop-wisely TBAF (85 mL, 85 mmol, 4.7 eq., TBAF 1 M in THF). Thereaction mixture was stirred at rt for 3 h, and then quenched with NH₄Clsat. (250 mL) and Et₂O (250 mL). The layers were separated and theaqueous was extracted with Et₂O (2×150 mL). The combined organic layerswere washed with brine (2×100 mL), dried with Na₂SO₄ and concentratedunder reduced pressure, purified on a SiO₂ column (75 g of 230-400 Meshsilica from Silicycle). The crude material was dissolved in CH₂Cl₂(15-20 mL) prior to be loaded on the column. The column was preparedusing 25% ethyl acetate/hexane. Elution: 25%, 35%, 50% ethylacetate/hexane. The desired material 8 (5.10 g, 50% combined yield from5) is a white foam.

Preparation of Intermediate for C3-C4 Modification Series:

To the stirred suspension of 2-deoxy-D-ribose (100.8 g, 0.75 mol,commercially available from TCI) in EtOAc (800 ml), were added2-methoxypropene (94 ml, 0.98 mol, 1.3 eq., Aldrich) and PPTS (4.16 g,17 mmol, 2 mol %, Aldrich) at room temperature under N₂.

The mixture was stirred vigorously for 3 hr.

Then the insoluble residue (remained SM) was filtrated out and TEA (4.6ml, 2 eq. to PPTS) was added to the filtrate. The resultant filtrate wasconcentrated under reduced pressure and the residue was purified bysilica gel flash column chromatography (silica gel 4 kg, hexane-EtOAc9:1 to 1:1 as eluent) to give 68 g of desired compound as colorless oil(52%).

To the stirred suspension of LiAlH₄ (9.26 g, 0.244 mol, 1.25 eq., Wako)in THF (200 mL) cooled in ice/brine bath, was added drop wise SM (68 g,0.39 mol) in THF (600 mL+100 mL rinse) under N₂ (in ca. 1.5 hr). Thenthe mixture was stirred for additional 15 min. After quenching bycareful addition of MeOH, 9.26 ml of water, 9.26 ml of 10% NaOH aq.,27.78 mL of water were added successively and the mixture was stirredvigorously for 1 hr. Then, the insoluble material was filtered out usingCelite and washed with EtOAc (500 mL×4), and the resultant filtrate wasconcentrated under reduced pressure, dried in vacuo to give 62.23 g ofthe crude product as light yellow oil (90.5%).

To the stirred suspension of NaH (8.09 g, 60% oil dispersion, 202 mmol,2.2 eq., Wako) in DMF (200 mL) cooled in ice/brine bath, was added dropwise (16.2 g, 91.9 mmol) in DMF (500 mL+100 mL rinse) under N₂. Theresultant mixture was stirred for 75 min. at room temperature. Then themixture was cooled to −55° C. (inner temp.)**, PivCl (12.5 mL, 102 mmol,1.1 eq., TCI) was added drop wise (in ca 10 min). After addition, themixture was allowed to warm to −30° C.

Quenching was performed by careful addition of sat. NH₄Cl aq., then themixture was extracted with EtOAc (1 L). After re-extraction of theaqueous layer with EtOAc (500 mL), the combined organic phase was washedwith water (0.6 L×3), brine (0.3 L) and dried over anhydrous Na₂SO₄.After filtration of drying agent, the filtrate was concentrated and theresidual brown oil (25.43 g) was purified by silica gel flash columnchromatography (silica gel 2.8 kg, hexane-EtOAc 2:1 to 1:1 as eluent) togive 3.73 g of less polar undesired protected mono-ol, 531-YW-2-2 (16%)and 13.14 g of polar desired product, 531-YW-2-3 (55%), respectively ascolorless oil.

Iodide Formation:

To a solution of 531-YW-2-3 (9.5 g, 36.5 mmol) in 400 mL of toluene,PPh₃ (18.3 g, 62.1 mmol, 1.9 eq.) was added. Then MeI (2.94 mL, 47 mmol,1.3 eq.) and DEAD (6.29 mL) were added simultaneously by twosyringe-pumps in 20 min. After stirred at room temp for 20 min, it waspoured into a rapid stirred pentane solution. The solid was dissolved bysmall amount of CH₂Cl₂ and added into the pentane. The precipitation wasfiltered through celite, the pad was washed with pentane. The combinedfiltrates were concentrated. The crude oil was purified quickly on ashort silica gel column with 20:1, 10:1, 6:1 Hex/EtOAc. It gave 11.6 gof the iodide 531-YW-3.

Coupling:

To a solution of iodide (531-YW-3, 11.6 g, 31.3 mmol) and selenide(509-HD-213, 24.5 g, 50.9 mmol, 1.6 eq.) in a mixed solvent of THF andHMPA (130 mL, 10:1 ratio), a solution of LiHMDS (94 mL, 0.5 M) was addedby a syringe pump in one and half hour at −78° C. After 20 min at −78°C., it was warmed to 0° C. After cooled back to −10° C., it was quenchedwith aq. sat. NH₄Cl and extracted with EtOAc (2×). The organic layerswere washed with brine, dried and concentrated. The crude product waspurified on silica gel with Hexanes/EtOAc, 20:1, 10:1, 6:1, 4:1, and 2:1to give 16.0 g.

The product from the above (16 g) was dissolved in CH₂Cl₂ (200 mL),MCPBA (16 g, 50.9 mmol, 1.6 eq., 55%) was added at 0° C. After 15 min at0° C., Triethylamine (20 mL, excess) was added. After 30 min, it wasquenched with aq. Sat. Na₂S₂O₃. After stirred for 20 min, it wasextracted with EtOAc (2×). The organic layers were washed with sat.Na₂S₂O₃, sat. NaHCO₃, brine, dried and concentrated. The crude productwas purified on silica gel with Hexanes/EtOAc, 20:1, 10:1, 3:1, gave12.5 g of the desired product, 531-YW-4 (83% in three steps).

To a solution of ester (5.66 g, 10 mmol) In EtOH (100 mL), 50 mL of 1NNaOH was added. The reaction was stirred at rt overnight. The reactionwas then diluted with water, extracted with EtOAc (3×). The combinedorganic layers were washed with brine, dried and concentrated. It waspurified on silica gel column to give 4.42 g of desired product as anoil (92%).

To a solution of (COCl)₂ (2.2 mL, 3 eq.) in 50 mL of CH₂Cl₂, DMSO wasadded slowly at −78° C. After 15 min at −78° C., a solution of alcohol(4.1 g, 3.5 mmol) was added into the reaction at −78° C. After 30 min atthat temperature, TEA (10.7 mL, 9 eq.) was added. The reaction waswarmed to rt. It was quenched with Sat. NH₄Cl, extracted with EtOAc(2×). The combined organic layers were washed with brine, dried andconcentrated to dryness. It was used in next reaction withoutpurification.

The aldehyde was used as a general intermediate for the synthesis ofC3-C4 modification by coupling with appropriate acetylene or equivalent.

Preparation of Acetylenes for C3-C4 Modifications:

To a solution of TMS-acetylene (38.8 mL) In 1 L of THF at −60° C.,n-BuLi (110 mL, 2.5 M) was added. The reaction was warmed to 0° C.briefly, then cooled back down to −60° C. BF₃.Et₂O (33.8 mL) was thenslowly added, followed by the epoxide (15 mL) via syringe pump. Afterstirred at −60° C. for 1.5 h, it was warmed to rt, quenched by Sat.NH₄Cl, extracted with EtOAc (2×). The organic layers were dried andconcentrated. The crude product was purified by silica gel column with4:1 Hexanes/EtOAc to give 13.9 g of desired product as an oil.

The alcohol was silylated under standard condition with TBSCl andImidazole in methylene chloride.

A mixture of TBS protected TMS-acetylene (8.7 g) and K₂CO₃ (8 g) inmethanol (120 mL) were stirred for 5 h at rt. It was extracted withEtOAc (2×). The organic layers were dried and concentrated. The crudeproduct was purified on silica gel with hexanes to give 6.17 g ofcolorless oil (95%)

The following acetylenes were prepared analogous to the preparationdescribed:

Preparation of ER803064:

Acetylene (2.65 g, 12.5 mmol) was dissolved in 30 mL of THF and cooleddown to −78° C. n-BuLi (2.5 N, 6.24 mL) in Hexane was added. Thereaction mixture was stirred at −78° C. for 10 min, and then a solutionof aldehyde (3.0 g, 6.24 mmol) in 30 mL of THF was added via cannula.The reaction mixture was stirred at −78° C. for 30 min and warmed upgradually to room temperature. It was quenched with water and extractedwith EtOAc. After purification on silica gel column, 3.7 g of 509-HD-108was obtained as a pale yellow oil in 78% yield.

509-HD-108 (3.4 g, 4.91 mmol) was dissolved in 200 mL hexane. Quinoline(200 μL) and Lindlar catalyst (500 mg) were added. The reaction mixturewas stirred at 40° C. under H₂ balloon atmosphere for a total of 18 h.Then the catalyst was filtered away. Quantitative amount of 509-HD-112was obtained as a pale yellow oil.

509-HD-112 (3.4 g, 4.9 mmol) was dissolved in 60 mL of dichloromethaneat room temperature. Triethylamine (1.71 mL, 9.8 mmol), benzoyl chloride(1.14 mL, 12.2 mmol) and catalytic amount of DAMP were added,respectively. After stirring for 12 h, 0.1N sodium hydroxide solutionwas added and the reaction mixture was extracted with EtOAc. The crudeproduct was purified on silica gel column, giving 509-HD-115 as acolorless oil in 94% yield.

509-HD-115 (3.7 g, 4.64 mmol) was dissolved in 50 mL of THF. The THFsolution of TBAF (1N, 25 mL) was added. The reaction mixture was heatedat 50° C. for 24 h. It was diluted with Et₂O and washed with H₂O. Afterpurification on silica gel column, 509-HD-116 was obtained as a paleyellow foam in 68% yield.

2-Chloro-1-methylpyridinium iodide (2.4 g, 9.5 mmol) and n-Bu₃N (2.3 mL,9.5 mmol) were dissolved in 180 mL of dichloromethane and heated toreflux. The solution of 509-HD-116 (1.85 g, 3.2 mmol) in 50 mL of THFwas added slowly. The reaction mixture was heated for 30 min. It waswashed with 0.02 N hydrochloric acid, sat. sodium bicarbonate solutionand brine, respectively. After purification on silica gel column,509-HD-118 was obtained as a pale yellow foam in 62% yield.

509-HD-118 (1.22 g, 2.2 mol) was dissolved in 30 mL of ethanol. Sodiumhydroxide (1N, 21.5 mL) solution was added. The reaction mixture wasstirred for 48 h at room temperature. It was diluted with H₂O, extractedwith EtOAc. After purification on silica gel column, 346 mg of the majordesired single isomer 509-HD-119B was obtained as a colorless oil.

509-HD-119B (155 mg, 0.34 mmol) was dissolved in 9 mL ofdichloromethane. Molecular sieve (4 A, 360 mg) and PCC (360 mg, 1.7mmol) were added. The reaction mixture was stirred for 1 h at roomtemperature. After passing through celite, 509-HD-125 was obtained ascolorless oil in quantitative yield.

509-HD-125 was dissolved in 2.5 mL of dichloromethane. Then hydrofluoricacid (6 N, 10 mL) was added. The reaction mixture was stirred at roomtemperature for 30 min. It was diluted with more dichloromethane, washedwith water and sat. sodium bicarbonate solution. After purification on aplug of silica gel, ER803064 was obtained as a white solid in 86% yield.

ER805149 were synthesized in similar manner starting from thecorresponding acetylene.

Preparation of B2526 (Trans Cyclopentane with Desired DiolConfiguration)

Step 1

To a solution of (trimethylsilyl)acetylene (8.8 mL, 62 mmol) in dry THF(100 mL), at −78° C. under an inert atmosphere, were added a 2.5Msolution of n-butyllithium (24.9 mL, 62 mmol) and boron trifluoridediethyl etherate (7.66 mL, 62 mmol). The reaction mixture was thentreated drop wise with a solution of cyclopentene oxide (2.71 mL, 31mmol). The reaction mixture was stirred at −78° C. for 1 hour thenwarmed to room temperature. The usual work up gave compound 453-MS-226(3.92 g; 69%).

Step 2

To a mixed solution of 453-MS-226 (3.77 g, 20.67 mmol) and4-methoxybenzyl 2,2,2-trichloroacetimidate (7 g, 24.8 mmol) in diethylether (80 mL), at room temperature under an inert atmosphere, was addeda 1M solution of trifluoromethane sulphonic acid in diethyl ether (0.62mL) drop wise over approximately 15 minutes. Two extra aliquots (1 mLeach) of the 1M trifluoromethane sulphonic acid solution were added at10 minutes and at 60 minutes. The usual work up, with subsequent partialpurification by chromatography, gave impure compound 453-MS-228 (3.37 g,approximately 54%), which was used in the next step without furtherpurification.

Step 3

A solution of impure 435-MS-228 (3.37 g, approximately 0.011 mol) inmethanol (33 mL) was treated with potassium carbonate (3.075 g, 0.022mol) and stirred for 3.5 hours. The usual work up, followed bychromatographic purification, gave compound 453-MS-230 (2.22 g,approximately 88%).

Step 4

To a solution of 453-MS-230 (1 g, 4.37 mmol) in dry THF (10 mL), at −78°C. under an inert atmosphere, was added drop wise a 1.6M solution ofn-butyllithium (2.73 mL, 4.37 mmol). The reaction mixture was stirred at−78° C. for 10 minutes, then warmed momentarily to 0° C., and cooled to−78° C. A solution of the 343-YW-277 (1.5 g, 3.63 mmol) in dry THF (15mL) was added drop wise. The reaction mixture was stirred at −78° C. for30 minutes then warmed to room temperature. The usual work up, followedby chromatographic purification, gave compound 453-MS-232 (1.90 g, 82%).

Step 5

A solution of compound 453-MS-232 (1.68 g, 2.61 mmol) in hexane (30 mL)was hydrogenated, at room temperature and pressure, in the presence ofLindlar's catalyst (168 mg) and quinoline (30 μL) for 20 hours.Filtration and concentration in vacuo gave compound 453-MS-237 (1.68 g,assumed quantitative) which was used in the next stage withoutpurification.

Step 6

To a solution of compound 453-MS-237 (1.61 g, 2.5 mmol) in drydichloromethane (20 mL) were added triethylamine (2.09 mL, 15 mmol),DMAP (30 mg, 0.25 mmol), and benzoyl chloride (0.58 mL, 5 mmol). Thereaction mixture was stirred at room temperature for 3 days then workedup in the usual manner. Chromatographic purification gave compound453-MS-240 (1.29 g, 69%).

Step 7

To a solution of compound 453-MS-240 (300 mg, 0.4 mmol) in THF (6 mL)was added TBAF (210 mg, 0.8 mmol). The reaction mixture was stirred atroom temperature for 2.5 hours then worked up in the usual manner togive compound 453-MS-244 (145 mg, 71%) (m/z: 533.2516 measured [M+23],533.2510 calculated).

Step 8

A mixed solution of compound 453-MS-244 (737 mg, 1.44 mmol) andtriphenylphosphine (682 mg, 2.6 mmol) in dry toluene (10 mL) was cooledto 0° C. under an inert atmosphere. A solution of dibenzylazidodicarboxylate (1.033 g, 3.46 mmol) in toluene (5 mL), and methyliodide (117 μl; 1.88 mmol) were added, separately and simultaneously, tothe reaction mixture over approximately 15 seconds. The reaction mixturewas stirred at 0° C. for 15 minutes then allowed to warm to roomtemperature. After 30 minutes at room temperature the reaction mixturewas worked up in the usual manner. Chromatographic purification gavecompound 453-MS-253 (550 mg, 61%).

Step 9

A mixture of compound 453-MS-253 (475 mg, 0.765 mmol) and compound554-RB-260 (519 mg, 0.86 mmol) was dissolved in a solution of 10% HMPAin THF (6 mL) and cooled to −78° C. under an inert atmosphere. A 0.5 Msolution of LiHMDS in THF (1.83 mL, 0.916 mmol) was then added drop wiseover approximately 30 minutes. The reaction mixture was stirred at −78°C. for 1 hour then treated with a 1M solution of LiHMDS in THF (0.916mL; 0.916 mmol). After 20 minutes the reaction mixture was warmed to 0°C. The intermediate crude product was worked up in the usual manner andpurified partially by chromatography. The intermediate was dissolved indichloromethane (8 mL) and cooled to 0° C. A solution of approximately65% meta-chloroperbenzoic acid (249 mg) in dichloromethane (2 mL) wasadded portion wise. After 30 minutes triethylamine (0.65 mL) was addedand the usual work up, followed by chromatographic purification, gavecompound 453-MS-262 (348 mg, 47%).

Step 10

To a vigorously stirred biphasic mixture of compound 453-MS-262 (440 mg,0.538 mmol), dichloromethane (20 mL) and water (10 mL), was added dropwise a solution of DDQ (147 mg, 0.646 mmol) in dichloromethane (15 mL).After 1 hour at room temperature the reaction mixture was worked up inthe usual manner. Chromatographic purification gave two fractions ofpartially resolved diastereoisomers: Fraction A (less polar): a mixtureof 3 co-eluted diastereoisomers—compound 453-MS-277A (190 mg); FractionB (more polar): a single diastereoisomer—compound 453-MS-277B (122 mg);(total yield: 312 mg, 83%)

Step 11

A solution of compound 453-MS-277B (122 mg, 0.175 mmol) intetrahydrofuran (5 mL) was treated with a solution of TBAF (92 mg, 0.35mmol) in THF (1 mL). The reaction mixture was stirred at roomtemperature for 6 hours then worked up in the usual manner to giveimpure compound 453-MS-279 (104 mg), which was used in the next stagewithout purification.

Step 12

A solution of crude compound 453-MS-279 (80 mg, assumed to contain 0.134mmol) in dichloromethane (30 mL) was treated with2-chloro-1-methylpyridinium iodide (45 mg, 0.174 mmol) andtri-n-butylamine (42 μL, 0.174 mmol). The reaction mixture was heatedunder reflux for 25 minutes then cooled to room temperature. The usualwork up and chromatographic purification gave compound 453-MS-284 (30mg, 39% from compound 453-MS-277B).

Step 13

A solution of compound 453-MS-284 (30 mg, 51 μmol) in a mixture ofethanol (1 mL) and tetrahydrofuran (0.5 mL) was treated with 1M aqueoussodium hydroxide (518 μL) and stirred for approximately 3 days at roomtemperature. The usual work up, followed by chromatographicpurification, gave compound 453-MS-289 (16 mg, 65%).

Step 14

A solution of compound 453-MS-289 (15 mg, 31.6 μmol) in dichloromethane(1.5 mL) was treated with PCC (81 mg, 0.375 mmol) in the presence ofpowdered 4 Å molecular sieves (81 mg). The reaction mixture was stirredvigorously for 70 minutes at room temperature. Basification with excesstriethylamine, followed by partial chromatographic purification gaveimpure compound 453-MS-296 (approximately 7 mg), which was used in thenext stage without further purification.

Step 15

A solution of impure compound 453-MS-296 (approximately 7 mg) in amixture of acetonitrile (1600 μL) and dichloromethane (400 μL) wastreated with 48% aqueous hydrofluoric acid (400 μL). After 25 minutesthe usual work up followed by chromatographic purification gave compoundB2526 (1.1 mg, approximately 6% from compound 453-MS-289).

Preparation of B2538 (Cis Cyclopentane with Desired Diol Configuration)

Step 1

To a solution of compound 453-MS-277A (190 mg, 0.273 mmol) intetrahydrofuran (7 mL) was added a solution of TBAF (143 mg, 0.545 mmol)in THF (2 mL). After 1 hour at room temperature the reaction mixture wastreated with additional TBAF (20 mg, 0.076 mmol). After a further 3hours the reaction mixture was worked up in the usual manner to giveimpure compound 453-MS-281 (186 mg), which was used in the next stagewithout purification.

Step 2

To a solution of triphenylphosphine (31.5 mg, 0.12 mmol) in drytetrahydrofuran (2.5 mL) was added diethyl azidodicarboxylate (19 mL;0.12 μmol), at room temperature under an inert atmosphere. A solution ofimpure compound 453-MS-281 (36 mg, 0.06 mmol) in dry tetrahydrofuran(2.5 mL) was added. After 90 minutes additional triphenylphosphine (31.5mg, 0.12 mmol) and diethyl azidodicarboxylate (19 ml, 0.12 mmol) wereadded. After a further 30 minutes the reaction mixture was worked up inthe usual manner. Chromatographic purification gave compound 501-MS-6(19 mg, 54% from compound 453-MS-277A).

Step 3

A solution of compound 501-MS-6 (19 mg, 32.8 μmol) in a mixture ofethanol (1 mL) and THF (0.5 mL) was treated with 1M aqueous NaOH (380μL) and stirred for approximately 17 hours at room temperature. Thereaction mixture was then heated to 100° C. for approximately 30minutes. The usual work up gave compound 501-MS-8 (15.5 mg,quantitative).

Step 4

A solution of compound 501-MS-8 (15.5 mg, 32 μmol) in dichloromethane(3.2 mL) was treated with PCC (85 mg, 0.39 mmol) in the presence ofpowdered 4 Å molecular sieves (85 mg). The reaction mixture was stirredvigorously for 2 hours at room temperature. Basification with excesstriethylamine, followed by chromatographic purification gave compound501-MS-11 (12.5 mg, 83%).

Step 5

A solution of compound 501-MS-11 (12 mg, 25 mmol) in a mixture ofacetonitrile (2400 μL) and dichloromethane (600 μL) was treated with 48%aqueous hydrofluoric acid (600 μL). After 1 hour the usual work upfollowed by chromatographic purification gave compound B2538 (4 mg, 41%)(m/z: 411.1 [M+23, 100%], 412.1 [35%]).

Preparation of B2522 (Trans Cyclopentane with Undesired DiolConfiguration)

Step 1

A solution of compound 453-MS-277A (190 mg, 0.273 mmol) in THF (7 mL)was treated with a solution of TBAF (143 mg, 0.545 mmol) in THF (2 mL).The reaction mixture was stirred at room temperature for 6 hours thenworked up in the usual manner to give impure compound 453-MS-281 (186mg), which was used in the next stage without purification.

Step 2

A solution of crude compound 453-MS-281 (150 mg, assumed to contain0.251 mmol) in dichloromethane (15 mL) was treated with2-chloro-1-methylpyridinium iodide (84 mg, 0.327 mmol) andtri-n-butylamine (78 μL, 0.327 mmol). The reaction mixture was heatedunder reflux for 40 minutes then treated with additional2-chloro-1-methylpyridinium iodide (84 mg, 0.327 mmol) andtri-n-butylamine (78 μL, 0.327 mmol). The reaction mixture was heatedunder reflux for a further 1 hour then cooled to room temperature. Theusual work up and chromatographic purification gave compound 453-MS-290(72 mg, 50% from compound 453-MS-277A).

Step 3

A solution of compound 453-MS-290 (72 mg, 0.124 mol) in a mixture ofethanol (2.4 mL) and THF (1.2 mL) was treated with 1M aqueous NaOH (1.24mL, 1.24 mmol) and stirred for approximately 4 days at room temperature.The usual work up, followed by chromatographic purification, gave threefractions of partially resolved compounds:

Fraction A (less polar): an unascertained mixture of diastereoisomers(10 mg);

Fraction B (more polar): a mixture of 2 diastereoisomers—compound453-MS-292B (46 mg);

Fraction C (most polar): single diastereoisomer of starting material453-MS-290 (2 mg);

(total yield: 56 mg; 95%)

Step 4

A solution of compound 453-MS-292B (20 mg, 42 μmol) in dichloromethane(2 mL) was treated with PCC (109 mg, 0.505 mmol) in the presence ofpowdered 4 Å molecular sieves (109 mg). The reaction mixture was stirredvigorously for 55 minutes at room temperature. Basification with excesstriethylamine, followed by chromatographic purification, gave compound453-MS-299 (17 mg, 86%).

Step 5

A solution of compound 453-MS-299 (19 mg, 0.04 mmol) in a mixture ofacetonitrile (4 mL) and dichloromethane (1 mL) was treated with 48%aqueous hydrofluoric acid (1 mL). After 35 minutes the usual work upfollowed by chromatographic purification gave compound B2522 (9 mg, 58%)(m/z: 411.1443 measured [M+23], 411.1420 calculated).

ER804018 and ER804019 (C4-F):

LDA (2.0 M, 36 mL) was added to the solution of ethyl 2-fluoropropanate(7.23 g) and acetaldehyde (13.5 mL) in ether (100 mL) at −78° C. Afterthe addition finished the reaction flask was kept in ice bath andgradually warm to room temperature. The reaction was quenched withaqueous ammonium chloride after overnight stirring. The aqueous phasewas extracted with ether and the combined organic phase was dried oversodium sulfate. The solvent was stripped off and the residue wasdistilled in vacuo to give 541-YJ-97 (4.22 g).

Chloro-t-butyldiphenylsilane was added to the mixture of 541-YJ-97 (4.22g) and imidazole (3.5 g) in methylene chloride (50 mL) and stirredovernight. Aqueous sodium bicarbonate was added to the reaction mixture.The aqueous phase was extracted with ether and the combined organicphase was dried over sodium sulfate. The solvent was stripped off andthe residue was purified with flush chromatograph (hexane/acetate 50/1)to give 541-YJ-99 (7.55 g).

DIBAL-H in toluene (1.0 M, 25 ml) was added to 541-YJ-99 in toluene (85mL) at 0° C. The reaction was quenched in one hour with methanol/waterand filtrated through Celite. The residue was purified by flashchromatograph to yield 541-YJ-101 (536 mg).

Dess-Martin periodinane was added to 541-YJ-101 (511 mg) in methylenechloride (15 ml) at room temperature. The mixture was diluted with etherin 40 minutes and filtrated through Celite. The filtrate wasconcentrated, and the residue was purified by prepTLC (hex/acetate 7/1)to give 541-Yj-105 (355 mg, 70%).

Triphenylphosphine (2.08 g) was added to carbon tetrabromide (1.31 g) inmethylene chloride at room temperature. After stirring for 40 minutes541-YJ-105 was added and stirred for 2 hours. The mixture wasconcentrated and filtrated through silica gel (hexane/acetate 7/1) toproduce 541-YJ-106 (452 mg, 89%).

n-Butyllithium (2.5 M, 0.74 mL) was added to 541-YJ-106 (450 mg) in THF(10 mL) at −78° C. After one hour 541-YJ-108 was added. The reaction waskept at 0° C. for one hour and warmed to room temperature before it wasquenched with aqueous ammonium chloride. The aqueous phase was extractedwith ether and the combined organic phase was dried over sodium sulfate.The solvent was stripped off and the residue was purified with TLC(hexane/acetate 4/1) to 541-YJ-109 (394 mg, 54%).

The suspension of Lindlar catalyst (420 mg) and 541-YJ-109 (390 mg) inhexane (8 mL) was charged with hydrogen and stirred overnight. Thesuspension was filtrated through Celite and rinsed with acetate. Thefiltrate was concentrated to give 541-YJ-111 (378 mg).

Benzoyl chloride (254 mg) and DMAP (catalytic amount) was added to thesolution of 541-YJ-111 (378 mg) and triethylamine (0.5 mL) in methylenechloride (7 mL) at room temperature. The mixture was kept stirringovernight and quenched with aqueous sodium bicarbonate. The aqueousphase was extracted with ether and the combined organic phase was driedover sodium sulfate. The solvent was stripped off and the residue waspurified with TLC (hexane/acetate 7/1) to 541-YJ-115 (419 mg).

The solution of 541-YJ-115 (419 mg) and TBAF (1.0 M, 2.2 mL) in THF (8mL) was kept stirring overnight, and then diluted with water. Theaqueous phase was extracted with ether and concentrated. The residue waspurified with TLC (methylene chloride/methanol 10/1) to give 541-YJ-116(194 mg).

The solution of 541-YJ-115 (419 mg) and TBAF (1.0 M, 2.2 mL) in THF (8mL) was kept stirring overnight, and then diluted with water. Theaqueous phase was extracted with ether and concentrated. The residue waspurified with TLC (methylene chloride/methanol 10/1) to give 541-YJ-116′(21 mg).

541-YJ-116 (21 mg) was added to the reflux of 2-chloro-1-methylpyridiumiodide (32 mg) and tributylamine (23 mg) in methylene chloride (4 mL).After 2 hours reflux the mixture was stirred overnight. The mixture wasdiluted with ether and washed with HCl (1.0 N) and water. The residuewas purified with TLC (hexane/acetate 1/1) to give 541-YJ-118-1 (7.7 mg)and 541-YJ-118-2 (8.4 mg).

PCC was added to 541-YJ-118-1 (7.7 mg) and MS 4 A suspension inmethylene chloride (2 mL) at room temperature. The mixture was stirredfor 3 hours and filtrated through silica gel. The silica gel was elutedwith acetate and concentrated to give 541-YJ-119 (3.3 mg).

PCC was added to 541-YJ-118-2 (8.4 mg) and MS 4 A suspension inmethylene chloride (2 mL) at room temperature. The mixture was stirredfor 3 hours and filtrated through silica gel. The silica gel was elutedwith acetate and concentrated to give 541-YJ-120 (3.0 mg).

Hydrofluoric acid (49%, 1 mL) was added to 541-YJ-119 (8.0 mg) inacetonitrile (3 mL) and stirred for 15 minutes. The mixture was dilutedwith water and extracted with methylene chloride. The organic phase wasconcentrated and purified with a short silica gel pad to produce541-YJ-126 (5.1 mg, ER-804018).

Hydrofluoric acid (49%, 1 mL) was added to 541-YJ-120 (6.0 mg) inacetonitrile (3 mL) and stirred for 15 minutes. The mixture was dilutedwith water and extracted with methylene chloride. The organic phase wasconcentrated and purified with a short silica gel pad to produce541-YJ-126 (3.1 mg, ER-804019).

Preparation of ER804142 and ER804143, C4-CF3 Analog:

To neat trifluoropropanic acid (10.0 g) was added oxalyl chloride (7.5mL) at room temperature and kept the mixture at 50° C. overnight. Thenbenzyl alcohol was added and kept stirring for 10 hours. The reactionwas quenched with aqueous sodium bicarbonate and extracted withchloroform. The solvent was stripped off and the residual was purifiedwith flush chromatograph (hexane/acetate 20/1) to afford 541-YJ-139(16.05 g, 94%).

LDA (1.5 M, 53.9 ml) was added to the solution of 541-YJ-139 (7.23 g)and acetaldehyde (20.6 ml) in THF (180 mL) at −78° C. After the additionfinished the reaction flask was kept in ice bath and gradually warm toroom temperature. The reaction was quenched with aqueous ammoniumchloride after overnight stirring. The aqueous phase was extracted withether and the combined organic phase was dried over sodium sulfate. Thesolvent was stripped off and the residue was purified with flushchromatograph (hexane/acetate 20/1 to 10/1) to afford 541-YJ-141 (10.26g, 53%).

Chloro-t-butyldiphenylsilane (12.2 ml) was added to the mixture of541-YJ-141 (10.26 g) and imidazole (10.7 g) in methylene chloride (200ml) and stirred overnight. Aqueous sodium bicarbonate was added to thereaction mixture. The aqueous phase was extracted with chloroform andthe combined organic phase was dried over sodium sulfate. The solventwas stripped off and the residue was purified with flush chromatograph(hexane/acetate 40/1) to give 541-YJ-143 (14.94 g, 76%).

DIBAL-H in toluene (1.0 M, 89.4 mL) was added to 541-YJ-143 in toluene(300 mL) at 0° C. The reaction was quenched in one and half hour withmethanol/water (70 mL/45 mL) and filtrated through Celite. The residuewas purified by flash chromatograph (hexane/acetate 10/1) to yield541-YJ-144 (7.38 g, 62%).

Dess-Martin periodinane (9.47 g) was added to 541-YJ-144 (7.38 g) inmethylene chloride (150 mL) at room temperature. The mixture was dilutedwith ether in one hour and filtrated through Celite. The filtrate wasconcentrated and the residue was purified by flash chromatograph(hexane/acetate 10/1) to yield 541-YJ-145 (7.37 g).

Triphenylphosphine (39.2 g) was added to carbon tetrabromide (24.74 g)in methylene chloride at room temperature. After stirring for 45 minutes541-YJ-145 (7.37 g) was added and stirred for 3 hours. The mixture wasconcentrated and purified by flash chromatograph (methylene chloride) toyield 541-YJ-146 (8.74 g, 85%).

n-Butyllithium (2.5 M, 3.47 mL) was added to 541-YJ-106 (2.39 g) in THF(20 mL) at −78° C. After one hour 343-YW-277 (0.98 g) was added at −78°C. The reaction was kept at 0° C. for one hour and then warmed to roomtemperature before it was quenched with aqueous ammonium chloride. Theaqueous phase was extracted with ether and the combined organic phasewas dried over sodium sulfate. The solvent was stripped off and theresidue was purified with flash chromatograph (hexane/acetate, 3/1) to541-YJ-148 (1.40 g, 65%).

The suspension of Rieke zinc (9.0 mL) was carefully added to 541-YJ-148(390 mg) in methanol-water (20 mL/2 mL) at room temperature. Thesuspension was heated at 70° C. for one and half hour. The mixture wasfiltrated through Celite and rinsed with acetate. The filtrate wasconcentrated to give 541-YJ-151 (1.09 g) without purification.

Benzoyl chloride (0.52 ml) and DMAP (catalytic amount) was added to thesolution of 541-YJ-151 (0.97 g) and triethylamine (1.24 mL) in methylenechloride (15 mL) at room temperature. The mixture was kept stirringovernight and quenched with aqueous sodium bicarbonate. The aqueousphase was extracted with chloroform and the combined organic phase wasdried over sodium sulfate. The solvent was stripped off and the residuewas purified with flash chromatograph (hexane/acetate 5/1) to 541-YJ-158(0.96 g, 88%).

The solution of 541-YJ-158 (0.96 g) and TBAF (1.0 M, 4.9 mL) in THF (20mL) was kept at 50° C. overnight and then diluted with water. Theaqueous phase was extracted with ether and concentrated. The residue waspurified with flash chromatograph (acetate) to give 541-YJ-159 (186 mg,30%).

541-YJ-159 (186 mg) was added to the reflux of 2-chloro-1-methylpyridiumiodide (223 mg) and tributylamine (162 mg) in methylene chloride (25mL). After 2 hours reflux the mixture was cooled down. The mixture wasdiluted with ether and washed with HCl (1.0 N) and water. The residuewas purified with TLC (hexane/acetate 1/1) to give 541-YJ-160 (169 mg).

The solution of 593-YJ-160 (169 mg) and sodium hydroxide (1.0 N, 0.5 mL)in ethanol (5 mL) was kept at 75° C. overnight. The mixture wasconcentrated and diluted with aqueous ammonium chloride. The aqueousphase was extracted with ether and the combined organic phase wasconcentrated. The residue was purified by TLC (hexane/acetate, 1/1) toyield 593-YJ-161 (39 mg, 28%).

Dess-Martin periodinane (50 mg) was added to 541-YJ-161 (39 mg) inmethylene chloride (2 mL) at room temperature. The mixture was stirredfor 3 hours and diluted with ether. The mixture was filtrated throughCelite and purified with TLC (hexane/acetate, 2/1) to give 541-YJ-168-1(6.7 mg) and 541-YJ-168-2 (5.3 mg).

Hydrofluoric acid (49%, 1 mL) was added to 541-YJ-168-1 (6.7 mg) or541-YJ-168-2 (5.3 mg) in acetonitrile (3 mL) and stirred for 15 minutes.The mixture was diluted with water and extracted with chloroform. Theorganic phase was concentrated and purified with TLC (hexane/acetate1/4) to produce 541-YJ-174 (1.5 mg, ER-804142) or 541-YJ-175 (0.5 mg,ER-804143).

Preparation of C4-Oxo Analogs, NF0675, NF0879, NF0880, and NF0905

Synthetic Procedure for NF0675

Methyl S-lactate (20.8 g, 0.2 mol) was dissolved in dry THF (500 mL),imidazole (17.7 g, 0.26 mol) was added and the mixture was cooled to 0°C. in ice/water bath. Then TBDPSCl (60.5 g, 0.22 mol) was added, themixture was allowed to warm slowly to rt and stirred overnight afterwhich a saturated solution of NaHCO₃ was added. The mixture wasextracted with EtOAc and the organic extract was washed with a saturatedsolution of NaHCO₃, water, brine, dried with anhydrous Na₂SO₄, filteredand concentrated.

The crude product (74.59 g) was dissolved in dry Et₂O (300 mL) and thesolution was cooled to 0° C. in ice/water bath. Then LiBH₄ (4.36 g, 0.2mol) was added portionwise, the mixture was allowed to warm slowly to rtand stirred for 2 days after which a saturated solution of NH₄Cl wasadded slowly. The mixture was extracted with EtOAc and the organicextract was washed with a saturated solution of NH₄Cl, water, brine,dried with anhydrous Na₂SO₄, filtered and concentrated. The crudeproduct was purified by chromatography on silica gel using 20%EtOAc/hexane to give 59.67 g (0.19 mol, 95% 2 steps) of the protectedcompound TM-01.

To a solution of oxalyl chloride (2.5 eq., 75 mmol, 6.54 mL) in CH₂Cl₂(60 mL), DMSO (5 eq., 150 mmol, 10.6 mL) was added at −78° C. After 15min at −78° C., a solution of alcohol TM-01 (30 mmol, 9.44 g) in CH₂Cl₂(100 mL) was added over a period of 40 min. After 30 min at −78° C.,Et₃N (6.5 eq., 195 mmol, 27.2 mL) was added and the reaction was warmedto −50° C. and stirred for 30 min. It was quenched with a saturatedsolution of NH₄Cl (100 mL), extracted with EtOAc. The organic layer wasdried with Na₂SO₄, filtered and concentrated. The crude aldehyde (9.55g) was dissolved in dry THF (100 mL) and cooled to −78° C. Then 0.5Msolution of propargyl magnesium bromide in dry THF (1.7 eq., 50 mmol,100 mL) was added dropwise over a period of 30 min and the reaction waswarmed to −10° C. It was quenched with a saturated solution of NH₄Cl,extracted with EtOAc. The organic layer was dried with Na₂SO₄, filteredand concentrated. The crude alcohol was purified by chromatography onsilica gel using 5% EtOAc/hexane to give 5.125 g (15.1 mmol, 50% 2steps) of the desired alcohol.

The alcohol (5.124 g, 15.1 mmol) was dissolved in CH₂Cl₂ (55 mL),diisopropylethylamine (15.84 mL, 90.8 mmol) was added and the mixturewas cooled to 0° C. in ice/water bath. Then chloromethyl methyl ether(3.45 mL, 45.4 mmol) was added and the mixture was allowed to warm tort. After 2 days, it was quenched with a saturated solution of NH₄Cl,extracted with EtOAc. The organic layer was dried with Na₂SO₄, filteredand concentrated. The crude product was purified by chromatography onsilica gel using 5% EtOAc/hexane to give 4.866 g (12.7 mmol, 84%) ofTM-02.

491-HAD-46 (620 mg, 2.4 mmol) was dissolved in dry DMF (10 mL),imidazole (243 mg, 3.6 mmol) was added and the mixture was cooled to 0°C. in ice/water bath. Then TBSCl (430 mg, 2.9 mmol) was added, themixture was allowed to warm slowly to rt and stirred for 30 min afterwhich a saturated solution of NaHCO₃ was added. The mixture wasextracted with EtOAc and the organic extract was washed with a saturatedsolution of NaHCO₃, water, brine, dried with anhydrous Na₂SO₄, filteredand concentrated. The crude product was purified by chromatography onsilica gel using 5% EtOAc/hexane to give 856 mg (2.3 mmol, 96%) of thesilyl ether.

To a suspension of LiAlH₄ (65 mg, 1.7 mmol) in dry THF (7 mL) was addeda solution of the silyl ether (856 mg, 2.3 mmol) in dry THF (13.5 mL) at0° C. The mixture was allowed to warm slowly to rt and stirred for 50min after which EtOAc and 1N HCl were added. The mixture was extractedwith EtOAc and the organic extract was washed with water, a saturatedsolution of NH₄Cl, water, brine, dried with anhydrous Na₂SO₄, filteredand concentrated. The crude product was purified by chromatography onsilica gel using 25% EtOAc/hexane to give 654 mg (2.3 mmol, 99%) of thealcohol.

The alcohol (654 mg, 2.3 mmol) was dissolved in dry CH₂Cl₂ (25 mL). ThenDess-Martin periodinane (1.67 g, 3.94 mmol) was added and stirred for 4hr after which a saturated solution of Na₂S₂O₃ and a saturated solutionof NaHCO₃ were added. The mixture was extracted with EtOAc and theorganic extract was washed with a saturated solution of NaHCO₃, water,brine, dried with anhydrous Na₂SO₄, filtered and concentrated to give648 mg (2.3 mmol, quant.) of TM-03.

TM-02 (2.2 eq., 5.0 mmol, 1.89 g) was dissolved in THF (20 mL) andcooled to −78° C., under nitrogen. Then, n-BuLi (1.6M/hexane, 2.0 eq.,4.5 mmol, 2.8 mL) was added and the reaction was stirred at −78° C. for60 min. Aldehyde TM-03 (2.3 mmol, 648 mg) dissolved in THF (8 mL) wasadded to the solution and stirred for 60 min at −78° C. The solution wasallowed to warm to rt and stirred for 1.5 hrs. The mixture was quenchedwith water, extracted with EtOAc and the organic extract was washed withbrine, dried with Na₂SO₄, filtered and concentrated. The residue waspurified by chromatography on silica gel using 15% EtOAc/hexane to give1.393 g (2.1 mmol, 92%) of TM-04.

TM-04 (2.1 mmol, 1.39 g) was dissolved in hexane (40 mL). Then,quinoline (27 mg) and 5% Pd—BaSO₄ on carbon (88 mg) were added. H₂balloon was mounted and the mixture was purged 5× with H₂. Reaction wasstirred under hydrogen. After 27 hrs, reaction was stopped, catalyst wasfiltered through celite and mixture was concentrated under reducedpressure. The crude product was purified by chromatography on silica gelusing 15% EtOAc/hexane to give 957 mg (1.4 mmol, 69%) of TM-05 as majorisomer and 267 mg (0.4 mmol, 19%) of the diastereomer of allylic hydroxyposition.

Using same procedure for 554-RB-242, TM-05 (954 mg, 1.4 mmol) wasconverted to TM-06 (913 mg, 1.2 mmol, 83%).

TM-06 (912 mg, 1.2 mmol) was dissolved in THF (23 mL). Then, acetic acid(0.084 mL, 1.5 mmol) and 1.0M solution of tetrabutylammonium fluoride inTHF (1.23 mL, 1.23 mmol) were added at rt. The mixture was stirredovernight after which a saturated solution of NH₄Cl was added. Themixture was extracted with EtOAc and the organic extract was washed witha saturated solution of NaHCO₃, water, brine, dried with anhydrousNa₂SO₄, filtered and concentrated. The crude product was purified bychromatography on silica gel using 30% EtOAc/hexane to EtOAc as eluentsto give 362 mg (0.55 mmol, 47%) of TM-07 and 212 mg (0.50 mmol, 43%) ofTM-08.

Using same procedure for 554-RB-260, TM-07 (359 mg, 0.54 mmol) wasconverted to TM-09 (374 mg, 0.48 mmol, 89%).

Using same procedure for 5, TM-09 (372 mg, 0.48 mmol) was converted toTM-10 (339 mg, 0.35 mmol, 72%).

To a stirred solution of TM-10 (172 mg, 0.18 mmol) in THF (2 mL) andEtOH (2 mL) was added 1N NaOH aq. (2 mL) at rt. After 2.5 hrs, it wasquenched by 1N HCl and extracted with EtOAc. The organic extract waswashed with water, brine, dried with anhydrous Na₂SO₄, filtered andconcentrated. The crude product was purified by chromatography on silicagel using 30% EtOAc/hexane to give 143 mg (0.16 mmol, 93%) of the allylalcohol.

The allyl alcohol (143 mg, 0.16 mmol) was dissolved in THF (3 mL). Then,1.0M solution of tetrabutylammonium fluoride in THF (0.49 mL, 0.49 mmol)was added at rt. The mixture was stirred for 3 hrs after which 1N HClwas added. The mixture was extracted with EtOAc and the organic extractwas washed with water, brine, dried with anhydrous Na₂SO₄, filtered andconcentrated. The crude product was purified by chromatography on silicagel using 10% MeOH/EtOAc to give 87 mg (0.16 mmol, quant.) of TM-11.

To a stirred solution of TM-11 (87 mg, 0.16 mmol) in THF (3 mL) wereadded triethylamine (0.029 mL, 0.2 mmol) and 2,4,6-trichlorobenzoylchloride (0.032 mL, 0.2 mmol) at rt. After 16 hrs, the reaction mixturewas diluted with toluene (80 mL) and added dropwise to a solution ofN,N-dimethylaminopyridine (498 mg, 4.1 mmol) in toluene (80 mL) over aperiod of 8 hrs under reflux. The resultant mixture was stirred for 15hrs under reflux. After concentration under reduced pressure, theresidue was dissolved in EtOAc and washed with 5% citric acid aq.,water, brine and dried over anhydrous Na₂SO₄, filtered and concentrated.The crude product was purified by chromatography on silica gel using 30%EtOAc/hexane to give 53 mg (0.10 mmol, 64%) of TM-12.

Using similar procedure for 509-HD-125, TM-12 (39.4 mg, 0.078 mmol) wasconverted to TM-13 (36.8 mg, 0.073 mmol, 94%).

To a stirred solution of TM-13 (12 mg, 0.024 mmol) in THF (1 mL)-H₂O(0.5 mL) was added trifluoroacetic acid (1 mL) at 0° C. The mixture wasthen allowed to warm to rt. After 3.5 hrs, the mixture was poured into asaturated solution of NaHCO₃ and extracted with EtOAc. The organicextract was washed with water, brine and dried over anhydrous Na₂SO₄,filtered and concentrated. The crude product was purified bychromatography on silica gel using 30% EtOAc/hexane to give 1.2 mg(0.0028 mmol, 12%) of NF0675.

Synthetic Procedure for NF0879 and NF0880

Using modified procedure for TM-02, TM-15 (10.56 g, 31.6 mmol) wasobtained from 10.4 g (0.1 mol) of methyl S-lactate.

Using same procedure for TM-04, TM-03 (3.64 g, 12 mmol) was converted toTM-16 (5.29 g, 8.5 mmol, 69%).

Using same procedure for TM-05, TM-16 (5.28 g, 8.5 mmol) was convertedto TM-17 (4.91 g, 7.9 mmol, 93%).

From 4.904 g (7.8 mmol) of TM-17, TM-19 (1.120 g, 1.8 mmol, 23% 2 steps)and TM-20 (2.218 g, 4.4 mmol, 57% 2 steps) were obtained by similarprocedure for TM-07.

Using similar procedure for TM-10, 1.104 g (1.8 mmol) of TM-19 wasconverted to TM-22 (955 mg, 1.03 mmol, 58% 4 steps).

Using similar procedure for TM-11, 955 mg (1.03 mmol) of TM-22 wasconverted to TM-23 (593 mg, 0.98 mmol, 95% 2 steps).

Using similar procedure for TM-12, 590 mg (0.98 mmol) of TM-23 wasconverted to TM-24 (438 mg, 0.75 mmol, 77%).

Using similar procedure for TM-13, 209 mg (0.36 mmol) of TM-24 wasconverted to TM-25 (186 mg, 0.32 mmol, 89%).

Using similar procedure for NF0675, 186 mg (0.32 mmol) of TM-25 wasconverted to NF0879 (72 mg, 0.14 mmol, 45%).

NF0879 (16 mg, 0.032 mmol) was dissolved in CH₂Cl₂ (3 mL), H₂O (0.3 mL)and DDQ (2 eq., 0.064 mmol, 14.9 mg) were added and the mixture wasstirred vigorously at rt for 3 hrs. The mixture was quenched with asaturated solution of NaHCO₃ and diluted with EtOAc. The organic layerwas separated and washed with a saturated solution of NaHCO₃, brine,dried with Na₂SO₄, filtered and concentrated. The crude residue waspurified by chromatography on silica gel using 5% MeOH/CHCl₃ to give 5mg (0.013 mmol, 41%) of NF0880.

Synthetic Procedure for NF0905

TM-20 (2.218 g, 4.4 mmol) was dissolved in CH₂Cl₂ (45 mL), imidazole(520 mg, 7.6 mmol) was added and the mixture was cooled to 0° C. inice/water bath. Then TBSCl (768 mg, 5.1 mmol) was added, the mixture wasallowed to warm slowly to rt and stirred for 1.5 hrs after which asaturated solution of NaHCO₃ was added. The mixture was extracted withEtOAc and the organic extract was washed with a saturated solution ofNaHCO₃, water, brine, dried with anhydrous Na₂SO₄, filtered andconcentrated to give 2.79 g of TM-26 as crude product.

TM-26 (2.79 g) was dissolved in CH₂Cl₂ (45 mL), triethylamine (1.85 mL,13.3 mmol) and N,N-dimethylaminopyridine (54 mg, 0.44 mmol) were addedand the mixture was cooled to 0° C. in ice/water bath. Then benzoylchloride (1.03 mL, 8.9 mmol) was added, the mixture was allowed to warmslowly to rt and stirred overnight after which a saturated solution ofNaHCO₃ was added. The mixture was extracted with EtOAc and the organicextract was washed with a saturated solution of NH₄Cl, 5% citric acidaq., water, and brine, dried with anhydrous Na₂SO₄, filtered andconcentrated to give 4.33 g of TM-27 as crude product.

TM-27 (4.33 g) was dissolved in CH₂Cl₂ (50 mL), H₂O (5 mL) and DDQ (1.28g, 5.5 mmol) were added and the mixture was stirred vigorously at rt for2 hrs. The mixture was quenched with a saturated solution of NaHCO₃ anddiluted with EtOAc. The organic layer was separated and washed with asaturated solution of NaHCO₃, brine, dried with Na₂SO₄, filtered andconcentrated. The crude residue was purified by chromatography on silicagel using 20% EtOAc/hexane to give 2.99 g of TM-28 with slightly amountof impurities.

TM-28 (2.99 g) was dissolved in CH₂Cl₂ (60 mL),2,6-di-tert-butyl-4-methyl-pyridine (3.34 g, 16 mmol) and methyltrifrate (1.5 mL, 13 mmol) were added and the mixture was stirred underreflux overnight. The mixture was quenched with a saturated solution ofNaHCO₃ and diluted with EtOAc. The organic layer was separated andwashed with a saturated solution of NaHCO₃, brine, dried with Na₂SO₄,filtered and concentrated.

The crude residue (6.25 g) was dissolved in THF (60 mL). Then, 1.0Msolution of tetrabutylammonium fluoride in THF (5.5 mL, 5.5 mmol) wasadded at rt. The mixture was stirred for 1.5 hrs after which a saturatedsolution of NH₄Cl was added. The mixture was extracted with EtOAc andthe organic extract was washed with water, brine, dried with anhydrousNa₂SO₄, filtered and concentrated. The crude product was purified bychromatography on silica gel using 30% EtOAc/hexane to give 362 mg (0.73mmol, 16% 5 steps) of TM-29.

Using similar procedure for TM-09, 359 mg (0.72 mmol) of TM-29 wasconverted to TM-30 (376 mg, 0.62 mmol, 86%).

Using similar procedure for TM-23, 371 mg (0.61 mmol) of TM-30 wasconverted to TM-31 (207 mg, 0.42 mmol, 69% 5 steps).

Using similar procedure for TM-12, 207 mg (0.42 mmol) of TM-31 wasconverted to TM-32 (206 mg, 0.42 mmol, quant.).

Using similar procedure for TM-13, 206 mg (0.42 mmol) of TM-32 wasconverted to TM-33 (170 mg, 0.36 mmol, 83%).

Using similar procedure for NF0675, 170 mg (0.36 mmol) of TM-33 wasconverted to NF0905 (50 mg, 0.13 mmol, 35%).

Preparation of Compound ER-804003 (C3 Trifluoromethyl)

Step 1

A solution of ethyl (R)-3-hydroxy-4,4,4-trifluorobutanoate (2 g, 10.7mmol) in DMF (20 mL) was treated with tert-butyldiphenylsilyl chloride(8.34 mL, 32.1 mmol) and imidazole (3.28 g, 35.3 mmol). The reactionmixture was heated at 70° C. under an inert atmosphere for 16 hours. Theusual work up followed by partial purification by chromatography gaveimpure compound 557-MS-4 (1 g, approximately 22%).

Step 2

A solution of impure compound 557-MS-4 (380 mg, assumed to contain 0.89mmol) in dichloromethane (5 mL) was cooled to −78° C. and treated with a1M solution of diisobutylaluminum hydride in dichloromethane (1.79 mL,1.79 mmol). The reaction mixture was allowed to warm to room temperatureand worked up in the usual manner. Partial purification bychromatography gave impure compound 557-MS-10 (which was used in thenext stage without further purification).

Step 3

To a solution of triphenylphosphine (910 mg, 3.47 mmol) in drydichloromethane (6 mL), at 0° C. under an inert atmosphere, was addeddrop wise a solution of carbon tetrabromide (575 mg, 1.73 mmol) in drydichloromethane (3 mL). After 10 minutes a mixed solution of impurecompound 557-MS-10 (assumed to contain 0.86 mmol) plus triethylamine(0.181 mL, 1.3 mmol) in dry dichloromethane (3 ml) was added drop wise.The reaction mixture was worked up in the usual manner to give compound557-MS-14 (331 mg, 72% from 557-MS-4).

Step 4

To a solution of compound 557-MS-14 (331 mg, 0.61 mmol) in dry THF (2.5mL), at −78° C. under an inert atmosphere, was added drop wise a 1.6Msolution of n-butyllithium in hexanes (0.771 mL, 1.23 mmol). Thereaction mixture was warmed momentarily to 0° C. then re-cooled to −78°C. A solution of compound 480-XYL-075 (247 mg, 0.51 mmol) in drytetrahydrofuran (2.5 mL) was added drop wise. Warming to 0° C. followedby the usual work up and chromatographic purification gave compound557-MS-19 (300 mg, 57%).

Step 5

A solution of compound 557-MS-19 (300 mg, 0.35 mmol) in hexane (10 mL)was hydrogenated, at room temperature and 1 atmosphere pressure, in thepresence of Lindlar's catalyst (60 mg) and quinoline (4 μL) for 40 hours(fresh Lindlar catalyst (120 mg) added after 2 hours). Filtration andconcentration in vacuo gave compound 557-MS-22 (302 mg) which was usedin the next stage without purification.

Step 6

A solution of crude compound 557-MS-22 (assumed to contain 0.35 mmol) in1,2-dichloroethane (10 mL) was treated with benzoyl chloride (0.122 mL,1.05 mmol), triethylamine (0.293 mL, 2.1 mmol) andN,N-4-dimethylaminopyridine (21 mg, 0.175 mmol). The usual work upfollowed by chromatographic purification gave compound 557-MS-26 (177mg, 53% from compound 557-MS-19).

Step 7

A solution of compound 557-MS-26 (177 mg, 0.183 mmol) in THF (3 mL) wastreated with a 1M solution TBAF in THF (1.83 mL). The usual work up gavecrude compound 557-MS-43, which was used in the next stage withoutfurther purification (m/z: 623.3 [M−1, 24%], 249.1 [100%]).

Step 8

A solution of crude compound 557-MS-43 (84 mg, 0.13 mmol) in1,2-dichloroethane (90 mL) was added slowly to a heated solution (80°C.) of 2-chloro-1-methylpyridinium iodide (210 mg, 0.81 mmol) andtri-n-butylamine (0.192 ml, 0.81 mmol) in dichloromethane (90 mL). Theusual work up and chromatographic purification gave compound 557-MS-70(15.7 mg, 20% from compound 557-MS-26).

Step 9

A solution of compound 557-MS-70 (15.7 mg, 25 μmol) in a mixture ofethanol (1.8 mL) and THF (0.9 mL) was treated with 1M aqueous NaOH (0.3mL) and stirred for approximately 16 hours at room temperature. Theusual work up gave compound 557-MS-74 (6 mg, 46%).

Step 10

A solution of compound 557-MS-74 (9 mg, 18 μmol) in dichloromethane (3mL) was treated with PCC (58 mg, 0.269 mmol) in the presence of powdered4 Å molecular sieves (58 mg). The reaction mixture was stirredvigorously for 90 minutes at mom temperature. Basification with excesstriethylamine, followed by partial chromatographic purification gaveimpure compound 557-MS-77, which was used in the next stage withoutfurther purification (m/z: 523.1 [M+23, 100%], 365.1 [22%]).

Step 11

A solution of impure compound 557-MS-77 (1 mg, assumed to contain 2μmol) in a mixture of acetonitrile (200 μl) and dichloromethane (50 μl)was treated with 48% aqueous hydrofluoric acid (50 μl). After 25 minutesthe usual work up followed by chromatographic purification gave compoundER-804003 (0.3 mg, approximately 4% from compound 557-MS-74).

Preparation of Compound ER-803924 (C4 Benzyl)

Step 1

To a solution of freshly prepared LDA (0.053 mmol) in dry THF (30 mL),at −78° C. under an inert atmosphere, was added drop wise a solution ofmethyl (S)-3-hydroxybutanoate (3 g, 0.025 mol) in dry THF (30 mL). After2 hours at −78° C. the reaction mixture was treated drop wise withbenzyl bromide (9.06 mL, 0.076 mol), then warmed to room temperature.The usual work up followed by chromatographic purification gave compound501-MS-226 (1.28 g, 24%).

Step 2

A solution of compound 501-MS-226 (1.28 g, 6.14 mmol) in DMF (10 mL) wastreated with tert-butyldiphenylsilyl chloride (1.76 mL. 6.75 mmol) andimidazole (461 mg, 6.75 mmol) then heated at 50° C. for 4 hours. Theusual work up followed by chromatographic purification gave compound501-MS-251 (1.87 g, 68%).

Step 3

To a solution of compound 501-MS-251 (1.37 g, 3.06 mmol) in drydichloromethane (50 mL), at −78° C. under an inert atmosphere, was addeda 1.5M solution of DIBAL-H in toluene (5.11 mL, 7.66 mmol). The reactionmixture was then warmed to 0° C. and stirred at 0° C. for 2 hours. Theusual work up followed by chromatographic purification gave compound501-MS-255 (1.06 g, 83%).

Step 4

To a solution of oxalyl chloride (0.314 mL, 3.6 mmol) in drydichloromethane (8 mL), at −78° C. under an inert atmosphere, was addeddrop wise a solution of dimethylsulfoxide (0.51 mL, 7.2 mmol) in drydichloromethane (4 mL). After 30 minutes a solution of compound501-MS-255 (1.369 g, 3.27 mmol) in dry dichloromethane (8 mL) was addeddrop wise. The reaction mixture was stirred at −78° C. for 1 hour thentreated with triethylamine (2.28 mL, 16.35 mmol) then warmed slowly toroom temperature. The usual work up gave compound 501-MS-257 (1.24 g,91%).

Step 5

To a solution of triphenylphosphine (2.57 g, 9.81 mmol) in drydichloromethane (16 mL), at 0° C. under an inert atmosphere, was addeddrop wise a solution of carbon tetrabromide (1.63 g, 4.91 mmol) in drydichloromethane (8 mL). A mixed solution of compound 501-MS-257 (1.24 g,2.97 mmol) and triethylamine (0.501 ml, 3.6 mmol) was added drop wiseand the reaction mixture warmed to room temperature. The usual work upfollowed by chromatographic purification gave compound 501-MS-261 (1.47g, 79% from 501-MS-255).

Step 6

To a solution of compound 501-MS-261 (1.47 g, 2.57 mmol) in dry THF (10mL), at −78° C. under an inert atmosphere, was added drop wise a 1.6Msolution of n-butyllithium in hexanes (3.21 mL, 5.14 mmol). The reactionmixture was warmed momentarily to 15° C. then re-cooled to −78° C. Asolution of compound 480-XYL-075 (950 mg, 1.976 mmol) in dry THF (10 mL)was added drop wise. Warming to 0° C. followed by the usual work up andchromatographic purification gave compound 501-MS-265 (1.11 g, 63%).

Step 7

A solution of compound 501-MS-265 (1.11 g, 1.24 mmol) in hexane (70 mL)was hydrogenated, at room temperature and 1 atmosphere pressure, in thepresence of Lindlar's catalyst (350 mg) and quinoline (70 μL), for 5hours. Filtration and concentration in vacuo gave compound 501-MS-267(1.13 g, assumed quantitative), which was used in the next stage withoutpurification.

Step 8

A solution of compound 501-MS-267 (160 mg, 0.178 mmol) in THF (1 mL) wastreated with a 1M solution of TBAF in THF (0.89 mL). After approximately15 hours the usual work up gave crude compound 501-MS-279 (81 mg,approximately 82%).

Step 9

A solution of compound 501-MS-279 (81 mg, 0.146 mmol) in1,2-dichloroethane (100 mL) was added slowly to a heated solution (85°C.) of 2-chloro-1-methylpyridinium iodide (223 mg, 0.87 mmol) andtri-n-butylamine (0.207 ml, 0.87 mmol) in dichloromethane (100 mL). Theusual work up and chromatographic purification gave compound 501-MS-282(15 mg, 19%) (m/z: 555.4 [M−1; 100%], 511.4 [28%]).

Step 10

A solution of compound 501-MS-282 (15 mg, 0.027 mmol) in dichloromethane(3 mL) was treated with PCC (72 mg, 0.334 mmol) in the presence ofpowdered 4 Å molecular sieves (72 mg). The reaction mixture was stirredvigorously for 90 minutes at room temperature. Basification with excesstriethylamine, followed by chromatographic purification gave compound501-MS-284 (12 mg, 80%).

Step 11

A solution of compound 501-MS-284 (9 mg, approximately 0.016 mmol) in amixture of acetonitrile (1.2 mL) and dichloromethane (300 μl) wastreated with 48% aqueous hydrofluoric acid (300 μL). After 20 minutesthe usual work up followed by chromatographic purification gave compoundER-803924 (7.5 mg, quantitative).

Preparation of C3-hydrogen with C4-Methyl Analogs:

Synthesis of ER-804035:

To the methyl (S)-(+)-3-hydroxy-2-methylpropionate (14.4 g, 0.121 mol)in DMF (26 mL) at 0° C. were added 4-dimethylaminopyridine (0.79 g,0.006 mol), imidazole (14.3 g, 0.210 mol) andt-butyldimethylchlorosilane (23.8 g, 0.158 mol). The mixture was stirredat 0° C. for 10 min then at room temperature overnight. The mixture waspartitioned between ether and saturated sodium bicarbonate solution. Thetwo layers were separated and the aqueous layer was extracted with etherthree times. The ether extracts were combined, dried with sodium sulfateand concentrated. The crude sample was chromatographed on a silica gelcolumn eluting with 5% ethyl acetate in hexanes to get 29.6 g (98%) ofproduct, 480-XYL-073 with satisfactory ¹H-NMR data.

The compound 480-XYL-073 (5.04 g, 21.7 mmol) was dissolved in methylenechloride (216 mL) and cooled to −78° C. To this was addeddiisobutylaluminium hydride solution in methylene chloride (1.0M, 22 mL)at a rate of 13.3 mL per hour via syringe pump down the inner side offlask wall. After completion of addition, the mixture was stirredadditional 30 min. The reaction was quenched slowly with methanol downthe wall and added some saturated aqueous solution of potassium sodiumtartrate. The mixture was warmed up to it, added additional potassiumsodium tartrate solution and stirred vigorously for 1 h. The layers wereseparated and aqueous layer was extracted two times with methylenechloride. The combined organic layers were dried with sodium sulfate,filtered and concentrated to get 4.36 g of crude material, 480-XYL-077.The ¹H-NMR data analysis showed the desired product aldehyde: startingmaterial:over-rudeced alcohol ratio was 3:1.33:1. This mixture wasdirectly used for the synthesis of 480-XYL-079.

The compound 480-XYL-079 was synthesized following the same procedure asthe synthesis of 554-RB-228.

DMSO (0.5 mL) in methylene chloride (24 mL) was cooled to −78° C. Tothis, was added oxalyl chloride solution in methylene chloride (2.0M,1.7 mL) and the mixture was stirred for 10 min at −78° C. The compound531-YW-005 (1.4 g, 2.9 mmol) in a solution of methylene chloride (3 mLand rinsed, 2×2 mL) was added via a cannula, and the mixture was stirredfor 10 min at −78° C. To this, was added triethylamine (2.5 mL) dropwise and the mixture was stirred for 1 hour at −78° C., was warmed to 0°C. The reaction was diluted with large amount of ether and washed oncewith saturated ammonium chloride solution and water (1:1), and withwater (3×). The ether layer was concentrated in vacuo and re-dissolvedin large amount of ether. This was washed two times with water and oncewith brine. The ether layer was concentrated, azeotroped with ethylacetate and toluene and dried under high vacuum to get 1.36 g (98%),480-XYL-075 which showed good purity by ¹H-NMR spectrum. This materialwas immediately used for the synthesis of 480-XYL-081.

The synthesis of 480-XYL-084 followed the same procedure as thesynthesis of 554-RB-240.

The compound 480-XYL-084 (855 mg, 1.21 mmol) was dissolved in a mixtureof methanol and water (5:1, 60 mL). A slurry solution of Rieke-zinc inTHF (8 mL) was added and the mixture was heated to reflux with stirringfor three hours. The mixture was filtered through a plug of celite andsilica gel, rising with ethyl acetate. The filtrate was concentrated,re-dissolved in methylene chloride and washed with saturated ammoniumchloride solution and then with saturated sodium bicarbonate solution.The aqueous phase was back extracted two times with methylene chlorideand once with ethyl acetate. The combined organic layers were dried withsodium sulfate and concentrated to get 944 mg of crude material,480-XYL-075, which showed satisfactory purity by ¹H-NMR spectrum anddirectly used for the synthesis of 480-XYL-092.

The synthesis of 480-XYL-092 was the same as the procedure of thesynthesis of 554-RB-242.

ER-804035

The synthesis of ER-804035 from 480-XYL-092 was followed the sameprocedure as for the synthesis of ER-803064.

Preparation of ER804022:

Oxalyl chloride (6.5 mL, 74.1 mmol) was dissolved in 150 mldichloromethane at −78° C. Methyl sulfoxide (10.5 mL, 148.2 mmol) wasadded. After 20 min, the solution of starting material (5.2 g, 24.7mmol) in 50 mL of dichloromethane was added at −78° C. After stirringfor 1 h at −78° C., triethylamine (31.0 mL, 222 mmol) was added and thereaction mixture was warmed up to room temperature. It was quenched withsat. ammonium chloride and extracted with ethyl acetate. Afterpurification on silica gel column, 509-HD-183 was obtained in 79% yield.

Triphenylphosphine (13.4 g, 51.2 mmol) was dissolved in 100 mLdichloromethane at 0° C. Carbon tetrabromide (8.5 g, 25.6 mmol) wasadded. After 15 min, the solution of 509-HD-183 (4.1 g, 19.7 mmol) andtriethylamine (2.8 mL, 19.7 mmol) in 50 ml of dichloromethane was added.After stirring for 30 min, the reaction mixture was triturated withpentane. After purification on silica gel column, 509-HD-184 wasobtained in 88% yield.

509-HD-184 (553 mg, 1.52 mmol) was dissolved in 10 mL of THF at −78° C.The solution of n-butyl lithium (2.5M, 1.33 mL) in hexane was added.After 15 min at −78° C., the solution of 531-HYW-5 in 5 ml of THF wasadded. After stirring for 30 min at −78° C., the reaction mixture waswarmed up to room temperature. It was quenched with water and extractedwith ethyl acetate. After purification on silica gel column, 509-HD-185was obtained in 95% yield.

509-HD-185 (750 mg, 1.09 mmol) was dissolved in 40 mL of hexane.Quinoline (50 μL) and Lindlar catalyst (120 mg) were added. The reactionmixture was stirred at mom temperature under H₂ balloon atmosphere for 5h. Then the catalyst was filtered away. Quantitative amount of509-HD-186 was obtained.

509-HD-186 (861 mg, 1.09 mmol) was dissolved in 15 mL of dichloromethaneat room temperature. Triethylamine (380 μL, 2.73 mmol), benzoyl chloride(253 μL, 2.18 mmol) and catalytic amount of DAMP were added,respectively. After stirring for 20 h, 0.1N sodium hydroxide solutionwas added and the reaction mixture was extracted with ethyl acetate. Thecrude product was purified on silica gel column, giving 509-HD-187 in95% yield.

509-HD-187 (813 mg, 1.03 mmol) was dissolved in a mixture of 10 mL ofdichloromethane and 5 mL of water. DDQ (234 mg, 1.03 mmol) was added.After stirring at room temperature for 1 h, the reaction mixture wasquenched with sat. sodium bicarbonate solution and extracted with ethylacetate. After purification on silica gel column, 509-HD-188 wasobtained in 48% yield.

509-HD-188 (313 mg, 0.47 mmol) was dissolved in 15 mL of dichloromethaneat 0° C. Triethylamine (130 μL, 0.94 mmol) and methanesulfonyl chloride(54 μL, 0.71 mmol) were added. After stirring for 20 min, the reactionmixture was quenched with sat. sodium bicarbonate and extracted withdichloromethane. After purification on silica gel column, 509-HD-189 wasobtained in 93% yield.

509-HD-189 (327 mg, 0.44 mmol) was dissolved in 10 mL of DMF. Sodiumazide (85 mg, 1.32 mmol) and catalytic amount of tetrabutylammoniumiodide were added. After stirring at 85° C. for 2 h, the reactionmixture was diluted with ethyl acetate and washed with water. Afterpurification on silica gel column, 509-HD-190 was obtained in 93% yield.

509-HD-190 (297 mg, 0.43 mmol) was dissolved in 10 mL of THF. Thesolution of TBAF (1N, 1.3 mL) was added. The reaction mixture wasstirred at room temperature for 1 h. It was diluted with Et₂O and washedwith H₂O. After purification on silica gel column, 509-HD-191 (215 mg)was obtained in quantitative yield.

Trimethylphosphine (1N, 1.5 mL) was dissolved in a mixture of 15 mL ofTHF and 5 mL of water at room temperature. 509-HD-191 (215 mg, 0.31mmol) was added. After stirring for 12 h, it was concentrated andazeotroped with toluene. The residue was re-dissolved in 50 mldichloromethane. EDC (593 mg, 3.1 mmol) was added. After stirring for 2h, it was diluted with water and extracted with dichloromethane. Afterpurification on HPTLC, 509-HD-197 was obtained in 30% yield.

509-HD-197 (51 mg, 0.092 mmol) was dissolved in 5 mL of ethanol. Sodiumhydroxide (1N, 0.92 mL) solution was added. The reaction mixture wasstirred for 48 h at room temperature. It was diluted with H₂O, extractedwith EtOAc. After purification on HPTLC, 11.5 mg of the major desiredsingle isomer 509-HD-198 was obtained as colorless oil.

509-HD-198 (10.0 mg, 0.022 mmol) was dissolved in 3 mL ofdichloromethane. Molecular sieve (4 A, 48 mg) and PCC (48 mg, 0.22 mmol)were added. The reaction mixture was stirred for 48 h at roomtemperature. After purification on prep TLC, 509-HD-200 was obtained in35% yield.

509-HD-200 (13 mg, 0.0079 mmol) was dissolved in 0.25 mL ofdichloromethane. Then hydrofluoric acid (6N, 1 mL) was added. Thereaction mixture was stirred at room temperature for 1 h. It was dilutedwith more dichloromethane, washed with water and sat. sodium bicarbonatesolution. After purification on a plug of silica gel, ER804022 wasobtained as a white solid in quantitative yield.

Preparation of ER-803027-00-01

447-JCH-245B

To a magnetically stirred solution of 531-YW-2-2 (3.5 g, 13.4 mmol) in15 mL of dry DMF cooled to −0° C. (ice/water; external thermometer) wasintroduced NaH (0.39 g, 16 mmol) followed by benzyl bromide (0.34 g, 20mmol). After 18 hours of stirring at room temperature the reactionmixture was cooled down at 0° C. and water was added. The reactionmixture was diluted with water and extracted with Ethyl ether. The crudewas purified on silica gel (Hexane/EtOAc: 90/10) to afford 447-JCH-245B(0.93 g, 20% yield).

447-JCH-268B

To a magnetically stirred solution of 447-JCH-245B (0.93 g, 2.7 mmol) in8 mL of ethanol 1 M NaOH aqueous solution (4 mL) was added. After 18hours of stirring at room temperature the reaction mixture was dilutedwith water and extracted with ethyl ether. The crude was purified onsilica gel (Hexane/EtOAc: 60/40) to afford 447-JCH-268B (0.47 g, 67%yield).

447-JCH-271B

Using a procedure analogous to that described for the synthesis ofintermediate 531-YW-3, 447-JCH-268B (0.450 g, 1.69 mmol) was reactedwith triphenylphosphine (0.887 g, 3.38 mmol), DEAD (0.32 mL, 2 mmol) andmethyl iodide (0.158 mL, 2.54 mmol) in toluene (16.9 mL) to afford447-JCH-271B (0.553 g, 86% yield).

447-JCH-273B

To a magnetically stirred solution of 509-HD-213 (0.553 g, 1.47 mmol)and 447-JCH-271B (1.06 g, 2.2 mmol) in 6 ml of a 10:1 ratio of HMPA/THFmixture at −78° C. (dry ice/acetone; internal thermometer) was slowly(syringe pump) introduced a 1 M solution of LiHMDS in THF (2.2 mL, 2.2mmol) diluted with 2.2 mL of the same mixture. After 30 min. at −78° C.,the reaction mixture was warmed-up to 0° C. The reaction was thenquenched by addition of a saturated aqueous solution of ammoniumChloride. The reaction mixture was diluted with water and extracted withEthyl ether. The crude was purified on silica gel (Hexane/EtOAc: 90/10to afford 447-JCH-273B (0.910 g, 84% yield).

447-JCH-275B

To a magnetically stirred solution of 447-JCH-273B (0.910 g, 1.25 mmol)in 24 mL of dichloromethane at 0° C. (ice/water; external thermometer),MCPBA (0.66 g, 3.8 mmol) was added. After 15 min. of stirring at 0° C.,triethylamine was added (1.5 mL) and the reaction mixture was warmed-upto room temperature. After 45 min of stirring at room temperature a 10%solution of sodium thiosulfate in a saturated aqueous solution of sodiumbicarbonate was added and the mixture was stirred for 30 minutes. Thereaction mixture was diluted with water and extracted with Ethyl ether.The crude was purified on silica gel (Hexane/EtOAc: 80/20 to afford447-JCH-275B (0.70 g, 98% yield).

447-JCH-277B

To a magnetically stirred solution of 447-JCH-275B (0.71 g, 1.2 mmol) in15 mL of methanol at room temperature was introduced a catalytic amountof Pd 10% on carbon. The reaction mixture was stirred 18 hours at roomtemperature and then filtered through a pad of celite. The crude waspurified on silica gel (Hexane/EtOAc: 70/30) to afford 447-JCH-277B(0.58 g, 99% yield).

447-JCH-280A

Using a procedure analogous to that described for the synthesis ofintermediate 480-XYL-075, 447-JCH-277B (0.6 g, 1.2 mmol) was reactedwith Dess-Martin reagent (0.763 g, 1.8 mmol), and sodium bicarbonate(0.38 g) in dichloromethane (48 ml) to afford 447-JCH-280A (0.602 g).

447-JCH-282B

Using a procedure analogous to that described for the synthesis ofER-803064 (stage 509-HD-108), 447-JCH-280A (0.6 g) was reacted withintermediate 343-YW-276 (0.32 g, 1.56 mmol) in THF (18 mL) to afford447-JCH-282B (0.6 g, 70% yield from 447-JCH-277B).

447-JCH-283B

Using a procedure analogous to that described for the synthesis ofER-803064 (stage 509-HD-112), 447-JCH-282B (0.6 g) hydrogenated usingLindlar catalyst to afford 447-JCH-283B (0.61 g).

447-JCH-285B

Using a procedure analogous to that described for the synthesis ofER-803064 (stage 509-HD-115), 447-JCH-283A (0.72 g) was reacted withbenzoyl chloride (0.37 mL, 2.63 mmol) to afford 447-JCH-285B (0.78 g,93%).

447-JCH-287B

To a magnetically stirred solution of 447-JCH-285B (0.68 g, 0.86 mmol)in a 2/1 dichloromethane/water mixture (26 mL) at room temperature, DDQ(0.17 g) was added. After 40 minutes of stirring at room temperature,the reaction mixture was diluted with ethyl acetate washed once with anaqueous solution of sodium hydroxide (0.1 N) and twice with water. Thecrude product was purified by flash chromatography eluting withhexane/ethyl acetate: (60/40) to afford 447-JCH-287B (0.52 g, 88%).

447-JCH-288A

Using a procedure analogous to that described for the synthesis ofER-803064 (stage 509-HD-116), 447-JCH-287B (0.58 g, 0.86 mmol) wasreacted with TBAF (0.68 g, 2.6 mmol) in THF (2.6 mL) to afford447-JCH-288A (0.47 g).

447-JCH-290B

To a magnetically stirred solution of 447-JCH-288A (0.31 g, 0.54 mmol)and triphenylphosphine (0.34 g, 2.17 mmol) in THF (43 mL) at roomtemperature, DEAD (0.57 g, 2.17 mmol) was added. After 1 hour ofstirring at room temperature, the reaction mixture was concentratedunder vacuum. The crude product was purified by flash chromatographyeluting with hexane/ethyl acetate: (70/30) to afford 447-JCH-290B (0.21g, 70%).

447-JCH-294B

Using a procedure analogous to that described for the synthesis ofER-803064 (stage 509-HD-119), 447-JCH-290B (0.21 g, 0.38 mmol) wasreacted with sodium hydroxide (1M solution, 1.9 mL, 1.9 mmol) in ethanol(5.7 mL) to afford 447-JCH-294B (0.128 g, 73%).

447-JCH-295B

To a magnetically stirred solution of 447-JCH-294B (0.07 g, 0.155 mmol)in dichloromethane (15 mL) at room temperature sodium bicarbonate (0.08g) and Dess-Martin reagent (165 mg, 0.388 mmol) were added. After 45minutes of stirring at room temperature, a 10% (w/w) thiosulfatesolution in saturated aqueous solution of sodium bicarbonate was added.The reaction mixture was diluted with water and extracted with ethylether. The crude product was purified by flash chromatography elutingwith n-hexane/ethyl acetate: (60/40) 447-JCH-295B (0.06 g, 88%).

447-JCH-296B/ER-803027

Using a procedure analogous to that described for the synthesis ofER-803064 (stage 509-HD-125), 447-JCH-295B (0.017 g, 0.038 mmol) wasreacted with HF (48%) (0.85 ml) in acetonitrile (3.4 mL) to afford447-JCH-296B/ER-803027 (0.006 g, 97%).

Preparation of B2329:

447-SG-089A

A mixture of 1,3-propanediol (15 g, 197 mmol), p-anisaldehydedimethylacetal (37 mL, 217 mmol) and p-toluene sulfonic acid (35 mg) wasstirred under slight vacuum for 6 hours at 35° C. in DMF (35.5 mL). Thereaction mixture was cooled to room temperature, and then a saturatedaqueous solution of sodium bicarbonate was added. The reaction mixturewas diluted with water and extracted with ethyl acetate to afford447-SG-089A (35.8 g). The crude was used directly into the next stepwithout purification.

447-SG-89B

To a magnetically stirred solution of 413-SG-89A (12.95 g, 66.67 mmol)in dichloromethane (225 mL) cooled to −5° C. (Ice/salt, internalthermometer), 1 M solution of DIBAL-H in toluene (100 mL, 100 mmol) wasadded. After 2 hours of stirring at room temperature, the reaction wasquenched by addition of methanol (100 mL). After 2 hours of vigorousstirring, 100 ml of a saturated aqueous solution of sodium sulfate wasadded. After 1 hour of stirring, the reaction mixture was diluted with100 mL of ethyl ether and stirred at room temperature for half an hour.The reaction mixture was filtered through a plug of celite and thesolvent was removed by evaporation. The crude product was purified byflash chromatography eluting with hexane/ethyl acetate (2:1 then 1/1) toafford 447-SG-89B (11.45 88% yield).

413-SG-106A

To a magnetically stirred solution of 413-SG-89B (2 g, 10.9 mmol) indichloromethane (225 mL) cooled to 0° C. (Ice/water, externalthermometer) DMSO (2.5 mL, 35.67 mmol) was added, followed by P₂O₅ (5.06g, 35.67 mmol). After one hours of stirring at room temperature thereaction was cooled down to 0° C. and triethylamine (7.1 mL, 50.95 ml)was added. After 45 minutes of stirring at room temperature. Thereaction mixture was diluted with water and extracted withdichloromethane. The solvent was removed by evaporation. The residue wastriturated with ether and the solid was filtered-off and wash withether. The solvent was removed by evaporation to afford 413-SG-106A (2.1g), the crude was used directly into the next step without purification.

413-SG-106B

Using a procedure analogous to that described for the synthesis ofintermediate 343-YW-276, 413-SG-106A (10.9 mmol) was reacted withtriphenylphosphine (7 g, 26.49 mmol), carbon tetrabromide (4.39 g, 13.25mmol) and triethylamine (1.4 mL, 10.9 mmol) in dichloromethane (12.6mL). The crude product was purified by flash chromatography eluting withpentane/dichloromethane (1/1) to afford 413-SG-106B (3.04 g, 85% yield).

413-SG-110B

Using a procedure analogous to that described for the synthesis ofintermediate 554-RB-240B, 413-SG-106B (1.66 g, 4.73 mmol) was reactedwith n-BuLi (2.5M in toluene, 4.2 mL, 10.41 mmol) in THF (32.5 mL) at−78° C. The resulting alkynyl lithium was then reacted with intermediate343-YW-277 (1.64 g, 3.97 mmol) in THF (12 mL) at −78° C. The crudeproduct was purified by flash chromatography eluting with hexane/ethylacetate (3/1) to afford 413-SG-110B (2.01 g, 86% yield).

413-SG-167A

Using a procedure analogous to that described for the synthesis ofintermediate 554-RB-241, 413-SG-110B (1.4 g, 2.27 mmol) was dissolved inhexane (32 mL) and hydrogenated using Lindlar catalyst to afford413-SG-167A (1.4 g).

413-SG-169B

Using a procedure analogous to that described for the synthesis of554-RB-242, 413-SG-167A (1.37 g, 2.27 mmol) was reacted with benzoylchloride (0.53 mL, 4.54 mmol), triethylamine (0.79 mL, 5.68 ml) and acatalytic amount of DMAP in dichloromethane (12 mL). The crude productwas purified by flash chromatography eluting with hexane/ethyl acetate(3/1) to afford 413-SG-169B (1.58 g, 99% yield).

413-SG-169B

413-SG-169B (1.58 g, 2.22 mmol) was reacted with TBAF (0.88 g, 3.34mmol) in THF (6 mL) at room temperature. The reaction mixture wasdiluted with water and extracted with ethyl ether. The crude product waspurified by flash chromatography eluting with hexane/ethyl acetate (5/1and then 1/1) to afford 413-SG-169B (1.02 g, 98% yield).

Using a procedure analogous to that described for the synthesis of554-RB-260, 413-SG-170B (1.02 g, 2.17 mmol) was reacted withtriphenylphosphine (0.97 g, 3.69 mmol), DEAD (0.36 mL, 2.28 mmol) andmethyl iodide (0.175 mL, 2.82 mmol) in toluene (19 mL). The crudeproduct was purified by flash chromatography eluting with hexane/ethylacetate (9/1 and then 5/1) to afford 413-SG-163B (1.12 g, 98% yield).

413-SG-174B

Using a procedure analogous to that described for the synthesis of531-YW-4, 413-SG-173B (1.12 g, 1.93 mmol) was reacted with intermediate509-HD-213 (1.2 g, 2.51 mmol) and LiHMDS (1M solution in THF, 2.3 mL,2.3 mmol) in a 10 to 1 THF/HMPA mixture (17.3 mL) to afford 413-SG-174A.413-SG-174A (crude) was reacted with MCPBA (0.61 g, 1.93 mmol) andtriethylamine (1.6 mL, 11.6 mmol). The crude product was purified byflash chromatography eluting with hexane/ethyl acetate (5/1 and then3/1) to afford 413-SG-174B (0.895 g, 45% yield).

413-SG-177B

Using a procedure analogous to that described for the synthesis of453-MS-262, 413-SG-174B (0.89 g, 1.14 mmol) was reacted with DDQ (0.31g, 1.37 mmol) in a 2/1-dichloromethane/water mixture (48 mL). The crudeproduct was purified by flash chromatography eluting with hexane/ethylacetate (5/1 and then 3/1) to afford 413-SG-177B (0.454 g, 61% yield).

413-SG-179B

Using a procedure analogous to that described for the synthesis ofER-803064 (stage 509-HD-116), 413-SG-177B (0.45 g, 0.691 mmol) wasreacted with TBAF (0.542 g, 2.07 mmol) in THF (2.2 ml). The crudeproduct was purified by flash chromatography withdichloromethane/methanol: (95/5) to afford 413-SG-179B (0.31 g, 79%yield).

413-SG-180B

413-SG-179B (0.31 g, 0.54 mmol) was reacted with triphenylphosphine(0.175 g, 0.658 mmol) and DEAD (0.105 ml, 0.658 mmol) in THF (43 ml).The crude product was purified by flash chromatography with hexane/ethylacetate: (3/1) to afford 413-SG-180B (0.2 g, 69% yield).

413-SG-182A

Using a procedure analogous to that described for the synthesis ofER-803064 (stage 509-HD-119), 413-SG-180B (0.18 g, 0.334 mmol) wasreacted with sodium hydroxide (1M solution, 1.7 mL, 1.7 mmol) in a 2/1mixture ethanol/THF (10 mL) to afford 413-SG-182A (0.16 g). The crudeproduct was used for the next step without further purification.

413-SG-188B

Using a procedure analogous to that described for the synthesis ofER-803064 (stage 509-HD-125), 413-SG-182A (0.14 g, 0.32 mmol) wasreacted with PCC (0.84 g, 3.87 mmol) in dichloromethane (34 mL) withmolecular sieves 4 Å (800 mg). The crude product was purified by flashchromatography with hexane/ethyl acetate: (70/30) to afford 413-SG-188B(0.07 g, 52% yield).

413-SG-93B

Using a procedure analogous to that described for the synthesis ofER-803064 (final step), 413-SG-188B (0.067 g, 0.157 mmol) was reactedwith HF (6 M solution in acetonitrile, 17.7 ml) in dichloromethane (3.1ml). The crude product was purified by flash chromatography withhexane/ethyl acetate: (60/40) to afford 413-SG-193B/B-2329 (0.05 g, 91%yield).

Preparation of B2395:

413-SG-184B

To a magnetically stirred solution of 413-SG-178B (0.194 g, 0.289 mmol)and triethylamine (0.08 mL, 0.578 mmol) in dry dichloromethane cooled to0° C. (ice/water; external thermometer) was introduced methanesulfonylchloride (0.034 ml, 0.434 mmol). After 1 hour of stirring at 0° C. asaturated aqueous solution of sodium bicarbonate was added. The reactionmixture was diluted with water and extracted with dichloromethane. Thecrude was purified on silica gel (Hexane/EtOAc: 1/1) to afford413-SG-184B (0.215 g, 99% yield).

413-SG-185B

A solution of 413-SG-184B (0.216 g, 0.288 mmol), sodium azide (0.028 g,0.432 mmol) and catalytic amount of tetra-butyl ammonium iodide in DMFwas magnetically stirred at 85° C. After 90 minutes the reaction mixturewas concentrated under vacuum. The crude was purified on silica gel(Hexane/EtOAc: 5/1) to afford 413-SG-185B (0.086 g, 93% yield)

413-SG-186A

Using a procedure analogous to that described for the synthesis ofER-803064 (stage 509-HD-116), 413-SG-185B (0.19 g, 0.27 mmol) wasreacted with TBAF (0.21 g, 0.8 mmol) in THF (1 ml) to afford 413-SG-186A(0.14 g). The crude product was used in the next step withoutpurification.

413-SG-217A

To a magnetically stirred solution of 413-SG-186A (0.092 g, 0.156 mmol)in THF/water 4/1 (1.5 mL) at room temperature was introducedtrimethylphosphine (0.78 mL, 0.778 mmol). After 18 hours of stirring atroom temperature the reaction mixture was concentrated under vacuum. Theresidue was diluted with water and extracted with dichloromethane toafford 413-SG-217A. The crude was dried and used in the next stepwithout purification.

To a magnetically stirred solution of 413-SG-217A (0.156 mmol) indichloromethane (0.2 mL) at room temperature was introduced EDC (0.10mg, 0.504 mmol). After 4 hours of stirring at room temperature thereaction mixture was Concentrated under vacuum. The crude was purifiedon silica gel (Hexane/EtOAc: 1/1) to afford 413-SG-217B (0.036 g, 42%yield).

413-SG-221A

Using a procedure analogous to that described for the synthesis ofER-803064 (stage 509-HD-119), 413-SG-217B (0.036 g, 0.065 mmol) wasreacted with sodium hydroxide (1M solution, 0.3 mL, 0.3 mmol) in a 2/1mixture of ethanol/THF (2 mL) to afford 413-SG-221A (0.031 g). The crudeproduct was used for the next step without further purification.

413-SG-226A

Using a procedure analogous to that described for the synthesis ofER-803027 (stage 447-JCH-295), 413-SG-221A (0.014 g, 0.031 mmol) wasreacted with Dess-Martin reagent (80 mg, 0.188 mmol) and 2.6-lutidine(0.036 mL, 0.313 mmol) in dichloromethane (2.1 mL). The crude waspurified on silica gel (dichloromethane/methanol: 98/2) to afford413-SG-226A (0.005 g, 36% yield).

413-SG-235B

Using a procedure analogous to that described for the synthesis ofER-803064 (final step), 413-SG-226AB (0.01 g, 0.022 mmol) was reactedwith HF (1.5 M solution in acetonitrile, 5 mL) in dichloromethane (2mL). The crude product was purified by flash chromatography eluting withn-hexane/ethyl acetate: (3/1) to afford 413-SG-235B (0.004 g, 50%yield).

Preparation of C8-Deoxy Analogs, NF0530, NF0531, NF0552 and NF0761

L-Dimethylmalate (50 g, 308.4 mmol) was dissolved in dry THF (308 mL)and cooled to 0° C. Then BH₃-Me₂S complex (10M, 1.1 eq., 34 mL, 0.34mol) was added dropwise and then the mixture was allowed to warm to rt.After stirred for 90 min then re-cooled to 0° C., NaBH₄ (0.05 eq., 15.4mmol, 583 mg) was added and stirred for additional 60 min. The reactionwas quenched with MeOH, and concentrated under reduced pressure. Thecrude residue was purified by chromatography on silica gel (AcOEt/MeOH)to afford MK-001 (26 g, 63%).

To a solution of MK-001 (10.0 g, 74.6 mmol) and p-anisaldehyde dimethylacetal (16.5 mL, 96.9 mmol) in 150 mL of dry CH₂Cl₂ was added DL-10-CSA(35 mg, 0.15 mmol) at 0° C., then the reaction mixture was allowed towarm to rt gradually. After 1 day 0.042 mL of Et₃N was added thenevaporated. The crude product was purified by chromatography on silicagel (Hexane/AcOEt: 5/1 to 3/1) to afford MK-002 (12.1 g, 64%).

MK-002 (12.1 g, 48.0 mmol) was dissolved in dry CH₂Cl₂-DME (240 mL-240mL) and cooled to −78° C. Then DIBAL-H in hexane (1.0 M, 50.4 mL, 50.4mmol) was added dropwise over 30 min and the mixture was stirred foradditional 100 min at −78° C. The reaction was quenched with MeOH (6 mL)then poured into a stirred solution of AcOEt and aqueous saturated Na/Ktartrate. The organic extract was washed with brine, dried over Na₂SO₄,filtered and concentrated to afford crude oil of MK-003 (11.84 g), whichwas used for next step without purification.

Ph₃PCH₃ ⁺Br⁻ (34.3 g, 96.0 mmol) was dissolved in dry THF (320 mL). Themixture was cooled to 0° C., and n⁻BuLi in hexane (1.6 M, 51.0 mL, 81.6mmol) was added slowly. After stirring for 120 min, a solution of thecrude MK-003 (11.84 g) in 50 mL of dry THF was added slowly. Thereaction was stirred for 30 min at 0° C. and followed by overnight atrt, then quenched with saturated aqueous solution of NH₄Cl. The mixturewas extracted with AcOEt, washed with brine, dried over MgSO₄, filteredand concentrated to give crude oil. The crude oil was diluted withEt₂O-hexane, the generated precipitate was filtered off, then thefiltrate was evaporated. The residual oil was purified by chromatographyon silica gel (Hexane/AcOEt: 20/1 to 8/1) to afford oil of MK-004 (4.25g, 40% 2 steps).

To a solution of MK-004 (4.05 g, 18.4 mmol) in 92 mL of dry THF wasslowly added BH₃-Me₂S (2.0 M in THF, 4.60 mL, 9.2 mmol) at 0° C., thenthe reaction mixture was stirred at 0° C. for 90 min after which it wasallowed to warm to it and stirred for 120 min. The solution wasre-cooled to 0° C., then treated with aqueous 3N—NaOH (28 mL) andaqueous 30%-H₂O₂ (28 mL) with vigorous stirring. The mixture wasextracted with Et₂O, washed with saturated aqueous solution of Na₂SO₃,dried over MgSO₄, filtered and concentrated to gave crude oil. The crudeoil was purified by chromatography on silica gel (Hexane/AcOEt: 1/1 to2/3) to afford oil of MK-005 (2.64 g, 60%).

Using similar procedure for the synthesis of the TBS-ether, intermediateof TM-03, from 491-HAD-46, MK-005 (2.64 g, 11.1 mmol) was converted tocrude MK-006 (3.91 g). It was used for next step without purification.

MK-006 (3.91 g) was dissolved in 74 mL of dry CH₂Cl₂ and cooled to −78°C. DIBAL-H in hexane (1.0 M, 5 eq., 55.3 mL, 55.3 mmol) was addeddropwise over 30 min, and the solution was stirred at −78° C. foradditional 60 min after which it was allowed to warm to 0° C. over 50min. The reaction was quenched with MeOH (7 mL) then poured into astirred solution of AcOEt and saturated aqueous Na/K tartrate. Theorganic extract was washed with brine, dried over Na₂SO₄, filtered andconcentrated to give a crude oil. The crude oil was purified bychromatography on silica gel (Hexane/AcOEt: 3/1) to afford an oil,MK-007 (3.19 g, 81% 2 steps).

To a solution of (COCl)₂ (3 eq., 2.24 mL, 25.6 mmol) in 84 mL of dryCH₂Cl₂, DMSO (6 eq., 3.64 mL, 51.3 mmol) was added slowly at −78° C.After 15 min at −78° C., a solution of MK-007 (3.03 g, 8.55 mmol) wasadded dropwise into the reaction at −78° C. After 30 min at thattemperature, Et₃N (9 eq., 10.7 mL, 76.9 mmol) was added slowly. Thereaction mixture was allowed to warm to −10° C. gradually. It wasquenched with saturated aqueous solution of NH₄Cl, extracted withAcOEt-hexane, washed with aqueous solution of KHSO₄ then brine, driedover Na₂SO₄, filtered and concentrated to gave crude oil of MK-008 (3.52g). It was used for next step without purification.

Using similar procedure for the synthesis of TM-04 from TM-03 and TM-02,MK-008 (3.52 g) was coupled with NY-22 (6.06 g) and then converted topure MK-009 (5.76 g, 100% 2 steps) as mixture of diastereomers onpropargylic position.

Using similar procedure for the synthesis of TM-05 from TM-04, MK-009(5.75 g) was converted to crude MK-010 (6.11 g) as mixture ofdiastereomers on allylic position.

Using similar procedure for the synthesis of 554-RB-242 from 554-RB-241,crude MK-010 (6.11 g) was converted to pure MK-011 (5.93 g, 89% 2 steps)as mixture of diastereomers on allylic position.

To a stirred solution of MK-011 (5.93, 7.59 mmol) in 76 mL of 99.5%EtOH, PPTS (0.15 eq., 286 mg, 1.14 mmol) was added at rt, then themixture was warmed to 45° C. After 1 day it was diluted with AcOEt, thenwashed with saturated aqueous solution of NaHCO₃ and brine, dried overMgSO₄, filtered and concentrated to give crude oil. The crude oil waspurified by chromatography on silica gel (Hexane/AcOEt: 2/1) to affordoil of MK-012 (4.94 g, 98%).

Using similar procedure for the synthesis of 554-RB-260 from 554-RB-244,MK-012 (4.20 g, 6.30 mmol) reacted with DIAD and methyl iodide in thepresence of Ph₃P to give pure MK-013 (4.74 g, 97%) as mixture ofdiastereomers on allylic position.

To a stirred mixture of diphenol (13.0 g, 66.3 mmol), MeOH (6.2 mL, 152mmol), and iPr₂EtN (13.9 mL, 79.5 mmol) in 110 mL of CH₃CN, TMSCHN₂ inhexane (2M, 38.1 mL, 76.2 mmol) was added dropwise over 80 min at rt,then stirred overnight. The reaction was quenched with 5% citric acidaqueous solution and extracted with AcOEt. The organic extract waswashed with saturated aqueous solution of NaHCO₃ and brine, dried overMgSO₄, filtered and concentrated. The crude product was purified bychromatography on silica gel (Hexane/AcOEt: 9/1) to afford crystals ofMK-014 (12.4 g, 89%).

Using similar procedure for the synthesis of 509-HD-209 from 509-HD-207,MK-014 (5.2 g, 25 mmol) was converted to pure MK-015 (6.3 g, 100%).

Using similar procedure for the synthesis of 509-HD-211 from 509-HD-209,MK-015 (6.0 g, 23.5 mmol) was converted to a mixture of MK-016 (5.3 g,55%) and inseparable diselenide (2.9 g, 22%)

Using similar procedure for the synthesis of compound 4 from compound 2and compound 3, MK-013 (1.5 g, 1.93 mmol) was coupled with mixture ofMK-016 and diselenide (including 2.90 mmol of MK-016) to afford crudeoil of MK-017 (4.3 g). It was used for next step without purification.

Using similar procedure for the synthesis of compound 5 from compound 4,crude MK-017 (4.3 g) was converted to MK-018 (1.60 g, 92% 3 steps) ascompound purified.

MK-018 (1.59 g, 1.76 mmol) was dissolved in 25 mL of THF. Then,tetrabutylammonium fluoride (TBAF) in THF (1M, 7.0 mL, 7.0 mmol) wasadded at rt. The mixture was stirred for 38 hrs before saturated aqueoussolution of NH₄Cl was added. The mixture was extracted with AcOEt andthe organic extract was washed with brine, dried over Na₂SO₄, filteredand concentrated. The crude product was purified by chromatography onsilica gel (hexane/AcOEt: 5/3 to 1/2) to afford oil of MK-019 (1.10 g,93%).

To a stirred solution of MK-019 (1.07 g, 1.61 mmol) in 20 mL of EtOH wasadded 32 mL of aqueous 1N—NaOH, then the mixture was warmed to 100° C.After 32 hrs, it was quenched with 32 mL of aqueous 1N—HCl and extractedwith AcOEt. The organic extract was washed with brine, dried overNa₂SO₄, filtered and concentrated. The crude product was purified bychromatography on silica gel (AcOEt/MeOH: 9/1) to afford oil of MK-020(860 mg, 100%).

Using similar procedure for the synthesis of TM-12 from TM-11, MK-020(860 mg, 1.61 mmol) was converted to mixture of MK-021 and MK-022 (402mg, 49% 2 steps; MK-021: MK-022=85:15).

To a stirred suspension of Dess-Martin periodinane (1.01 g, 2.38 mmol)in 40 mL of dry CH₂Cl₂, a solution of MK-021 and MK-022 (402 mg, 0.794mmol) in 40 mL of dry CH₂Cl₂ was added at 0° C., then the mixture waswarmed to rt. After 14 hrs it was re-cooled to 0° C., diluted withAcOEt, washed with saturated aqueous solution of Na₂SO₃, NaHCO₃ andbrine, dried over Na₂SO₄, filtered and concentrated to give crude oil.The crude oil was purified by chromatography on silica gel(Hexane/AcOEt: 3/1) to afford colorless crystals of NF0552 (301 mg, 74%)and colorless oil of NF0530 (35 mg, 9%).

Using similar procedure for the synthesis of NF0675 from TM-13, NF0552(263 mg, 0.515 mmol) was converted to NF0530 (199 mg, 83%) as compoundpurified.

To a stirred mixture of NF0530 (233 mg, 0.499 mmol) in 17 mL of CH₂Cl₂and 1.7 mL of aqueous phosphate buffer (pH 6.86) was portionwise addedDDQ (283 mg, 1.25 mmol) at 0° C., then the mixture was allowed to warmto rt slowly. After 3.5 hrs it was quenched with aqueous solution ofNaHCO₃ and diluted with AcOEt. The organic extract was washed withaqueous solution of NaHCO₃ and brine, dried over Na₂SO₄, filtered andconcentrated. The crude product was purified by chromatography on silicagel (hexane/AcOEt: 5/2) to afford colorless crystals of NF0531 (143 mg,83%).

NF0552 (30 mg, 0.059 mmol) was treated with DDQ (3 eq.) at rt usingsimilar procedure for the synthesis of NF0531 from NF0530. The messyreaction was worked up in the usual manner. Purification bychromatography on silica gel (Hexane/AcOEt: 1/3) gave colorless oil ofNF0761 (1.7 mg, 7%). NF0761 was analyzed by HRMS; FAB+ m/z 407 (MH⁺),Anal. Calcd for C21H26O8: MH+, 407.1706. Found 407.1711 (MH+).

Preparation of C11-C12, cyclopropyl analogs, NF1226 and NF1227

Using the same procedure for the synthesis of TM-03 from 531-yw-2-3(491-HAD-46), MK-023 was obtained.

To a stirred mixture of MK-023 (2.5 g, 8.61 mmol) and 4-MPMCl (1.63 mL,12.0 mmol) in 40 mL of DMF, NaH (66%, 344 mg, 9.47 mmol) was addedportionwise at 0° C. and the mixture was warmed to rt. After stirred for3 hrs, the reaction was quenched with saturated aqueous solution ofNH₄Cl and extracted with AcOEt. The organic extract was washed withsaturated aqueous solution of NaHCO₃ and brine, dried over Na₂SO₄,filtered and concentrated. The crude product was purified bychromatography on silica gel (Hexane/AcOEt: 15/1) to afford colorlessoil of the fully protected tetraol (2.83 g, 80%).

The fully protected tetraol (3.11 g, 7.58 mmol) was dissolved in 38 mLof THF. Then, tetrabutylammonium fluoride (TBAF) in THF (1M, 9.9 mL, 9.9mmol) was added at rt. The mixture was stirred for 2 hrs beforesaturated aqueous solution of NH₄Cl was added. The mixture was extractedwith AcOEt and the organic extract was washed with brine, dried overNa₂SO₄, filtered and concentrated. The crude product was purified bychromatography on silica gel (Hexane/AcOEt: 1/1) to afford oil of MK-024(2.22 g, 99%).

Using similar procedure for the synthesis of 531-YW-3 from 531-YW-2-3,MK-024 (1.90 g, 6.41 mmol) was converted to MK-025 (2.36 g, 90%) ascompound purified.

Using similar procedure for the synthesis of 531-YW-4, MK-025 (2.36 g,5.80 mmol) coupled with 509-HD-213 (3.63 g, 7.54 mmol) was converted toMK-026 (3.10 g, 89% 3 steps) as compound purified.

To a stirred solution of MK-026 (2.0 g, 3.32 mmol) in 130 mL of toluenewere added Et₂Zn in hexane (1M, 16.6 mL, 16.6 mmol) and CH₂I₂ (1.34 mL,16.6 mmol) at −30° C. After stirred for 30 min at −30° C., it was warmedto it gradually over 2 hrs and then quenched with saturated aqueoussolution of NH₄Cl. (Note: To avoid decomposition of target product,short reaction time, 1-2 hrs, was required regardless of conversionrate.). The mixture was extracted with AcOEt and the organic extract waswashed with brine, dried over Na₂SO₄, filtered and concentrated. Thecrude product was purified by chromatography on silica gel(Hexane/AcOEt: 6/1 to 5/1) to afford oil of MK-027 (369 mg, <19%)including small amount of MK-026, and MK-026 (519 mg, <26%) includingsmall amount of MK-027 was recovered. The recovered MK-026 was treatedagain in the same manner to give MK-027 (367 mg, <29%) including smallamount of MK-026. Trans cyclopropane MK-027 was obtained as mixture ofinseparable stereoisomers (1:1).

MK-027 (736 mg, ca 1.19 mmol) including 10% of MK-026 was dissolved in24 mL of THF and 4 mL of water. OsO₄ in ^(t)BuOH (3 w/v %, 0.05 eq.,0.51 mL, 0.06 mmol), N-methylmorpholine (0.2 eq., 0.026 mL, 0.24 mmol)and NaClO₃ (0.4 eq., 51 mg, 0.48 mmol) were added into the stirredsolution at rt. After 2 days, to the mixture were added Celite, AcOEt,and aqueous solution of Na₂SO₃. The suspension was filtered, and thefiltrate was washed with brine, dried over Na₂SO₄, filtered andconcentrated to give crude oil of MK-027 (768 mg) which included noMK-026. It was used for next step without purification.

Using similar procedure for the synthesis of NF0531 from NF0530, thecrude oil of MK-027 (768 mg) was converted to MK-028 (523 mg, 88% 2steps) as compound purified.

Using similar procedure for the synthesis of MK-009 from MK-007, MK-028(204 mg, 0.411 mmol) was converted to MK-029 (231 mg, 69% 2 steps) ascompound purified.

MK-029 (231 mg, 0.283 mmol) was dissolved in 7 mL of 99.5% EtOH and 7 mLof hexane (note: no reaction in hexane). Then, quinoline (0.3 eq., 0.01mL, 0.085 mmol) and 5% Pd—BaSO₄ on carbon (0.05 eq., 30 mg, 0.014 mmol)were added. H₂ balloon was mounted and the mixture was purged with H₂.After stirred for 3.5 hrs under H₂ (1 atm) at rt, the reaction mixturewas filtered through Celite and the filtrate was evaporated to give acrude oil of cis-olefin (240 mg).

Using similar procedure for the synthesis of 554-RB-242 from 554-RB-241,the crude cis-olefin (240 mg) was converted to MK-030 (216 mg, 83% 2steps) as compound purified.

Using similar procedure for the synthesis of MK-019 from MK-018, MK-030(215 mg, 0.233 mmol) was directly converted to a carboxylic acid MK-031(125 mg, 92%) as compound purified.

Using similar procedure for the synthesis of TM-12 from TM-11, MK-031(123 mg, 0.210 mmol) was converted to crude lactonized product. It waspurified by chromatography on silica gel (Hexane/AcOEt: 4/1, 3/1, to1/1) to afford oil of MK-032 (27 mg, 22%) and oil of des-MOM form MK-033(20 mg, 18%).

To a stirred solution of MK-032 (24 mg, 0.0424 mmol) in 0.85 mL of EtOHand 0.85 mL of THF was added aqueous 1N NaOH (2.1 eq., 0.089 mL, 0.0889mmol). After stirred for 3 days at rt, the mixture was diluted withAcOEt and washed with brine, dried over Na₂SO₄, filtered andconcentrated to give crude product of MK-034 (24 mg). It was used fornext step without purification.

The crude MK-034 (24 mg, assumed to contain 0.0424 mmol) was dissolvedin 9 mL of CH₂Cl₂. To the solution were added Molecular sieve 4A (45 mg)and PDC (3 eq., 48 mg, 0.127 mmol) at rt. The reaction mixture wasstirred for 4 days at rt, then diluted with Et₂O and passed through apad of Celite. The filtrate was evaporated to give crude product. It waspurified by chromatography on silica gel (Hexane/AcOEt: 3/1 to 2/1) toafford colorless oil of MK-035 (less polar single isomer on transcyclopropane, 3.6 mg, 18% 2 steps), colorless oil of MK-036 (polarsingle isomer on trans cyclopropane, 5.8 mg, 30% 2 steps) and paleyellow oil of MK-037 (isomerized trans olefin, 5.4 mg, 28% 2 steps).

50% Hydrofluoric acid (24N, 0.2 mL) was added to less polar isomerMK-035 (3.6 mg, 0.00782 mmol) in 0.8 mL of CH₃CN and stirred for 1 hr at0° C. After stirred at rt for additional 1 hr, the reaction mixture wasquenched with saturated aqueous solution of NaHCO₃ and extracted withAcOEt. The organic extract was washed with brine, dried over MgSO₄,filtered and concentrated to crude product. It was purified bychromatography on silica gel (hexane/AcOEt: 1/2) to afford colorlesscrystals of NF1226 (2.3 mg, 59%) as single isomer.

Using similar procedure for the synthesis of MK-034 from MK-032, MK-033(20 mg, 0.0383 mmol) was converted to diol intermediate (13 mg, 81%).

Using similar procedure for the synthesis of NF530 from MK-022, the diolintermediate (15 mg, 0.0358 mmol) was converted to MK-038 (4.7 mg, 32%).

Using similar procedure for the synthesis of NF0675 from TM-13, MK-038(4.8 mg, 0.0115 mmol) was converted to colorless crystals of NF1227 (3.6mg, 83%, single isomer).

Using similar procedure for the synthesis of NF1226 from MK-035, polarisomer MK-036 (5.8 mg, 0.0126 mmol) was converted to colorless crystalsof NF1227 (2.6 mg, 55%, single isomer).

NF1227 differs from NF1226 as to stereochemistry on trans cyclopropane.

Preparation of C11-C12 Amide Analogs, NF1535, NF1537, and NF2306

Exemplary Synthetic Procedure for NF1535 and NF1537

Using similar procedure for the synthesis of NY-07 from NY-06, MK-014(12.40 g, 59.0 mmol) was converted to MK-039 (13.68 g, 92%) andpurified.

MK-039 (13.68 g, 54.2 mmol) was dissolved in 360 mL of CCl₄ and thesolution was heated up to reflux. To the stirred solution wasportionwise (over 1.5 hrs) added a mixture of NBS (11.1 g, 62.4 mmol,1.15 eq.) and (PhCO)₂O₂ (722 mg, 2.98 mmol, 0.055 eq.) and stirred foradditional 30 min at reflux. The reaction mixture was cooled to rt andinsoluble material was filtered off, then the filtrate was concentratedto give crude product. The crude product was purified by chromatographyon silica gel (hexane/AcOEt: 4/1 to 3/1) to afford colorless oil ofMK-040 (14.51 g, <81%) including small amount of starting material anddibromide. It was used for next step without further purification.

To a solution of MK-040 (14.51 g, assumed to contain 43.82 mmol) in 220mL of DMSO was added a solution of AgBF₄ (11.09 g, 57.0 mmol) in 55 mLof DMSO at rt. After 2 hrs, Et₃N (18.3 mL, 131.4 mmol) was added andstirred for 40 min at rt. The reaction mixture was diluted with AcOEt,then washed with saturated aqueous solution of NaHCO₃ and brine, driedover Na₂SO₄, filtered and concentrated to give a crude oil. The crudeproduct was purified by chromatography on silica gel (hexane/AcOEt: 3/1to 1/1) to afford colorless oil of MK-041 (5.49 g, 38% 2 steps).

To a stirred mixture of MK-041 (5.49 g, 20.63 mmol) in 140 mL of 99.5%EtOH was added imidazole (421 mg, 6.19 mmol) at rt. After stirred for 6days, the mixture was evaporated, diluted with AcOEt, washed with waterthen brine, dried over Na₂SO₄, filtered and concentrated to give crudecolorless crystals of MK-042 (4.42 g, <96%). It was used for next stepwithout purification.

To a stirred suspension of crude MK-042 (2.41 g, assumed to contain10.75 mmol) and K₂CO₃ (3.94 g, 28.5 mmol) in 70 mL of DMF was addedMOMCl (1.77 mL, 23.3 mmol) at 0° C., then the mixture was allowed towarm to rt. After 14 hrs, the reaction mixture was quenched withsaturated aqueous solution of NaHCO₃ and extracted with AcOEt. Theorganic extract was washed with brine, dried over Na₂SO₄, filtered andconcentrated to give crude oil of MK-043 (2.93 g, quant.). It was usedfor next step without purification.

Crude MK-043 (2.93 g, assumed to contain 10.75 mmol) was dissolved in 80mL of t-BuOH and 20 mL of water. Then 2-methyl-2-butene (5.69 mL, 53.7mmol, 5 eq.) and NaH₂PO₄-2H₂O (1.68 g, 10.75 mmol) were added. To thestirred suspension was portionwise added NaClO₂ (1.94 g, 21.5 mmol, 2eq.) at rt. After 1 hr at rt, the mixture was diluted with AcOEt andwater, then acidified with aqueous KHSO₄ solution to approximately pH 4.The organic extract was washed with brine, dried over Na₂SO₄, filteredand concentrated to give crude oil of MK-044 (3.20 g, quant.). It wasused for next step without purification.

Crude MK-044 (3.20 g, assumed to contain 10.75 mmol) was dissolved in 72mL of dry THF and 4.45 mL of BnOH (43.00 mmol, 4 eq.). Then Et₃N (1.80mL, 12.90 mmol, 1.2 eq.) and DPPA (2.54 mL, 11.82 mmol, 1.1 eq.) wereadded. The mixture was heated to 65° C. and stirred for 15 hrs, thencooled to rt. The mixture was diluted with AcOEt and saturated aqueoussolution of NH₄Cl. The organic extract was washed with saturated aqueoussolution of NaHCO₃ then brine, dried over Na₂SO₄, filtered andconcentrated to give a crude oil. The crude product was purified bychromatography on silica gel (hexane/AcOEt: 5/1) to afford colorlesscrystals of MK-045 (3.50 g, 80% 4 steps).

To a stirred solution of MK-045 (2.73 g, 7.02 mmol) in 80 mL of EtOH wasadded aqueous 0.5N—NaOH (1.15 eq., 16.2 mL, 8.07 mmol). After stirredfor 2 days at rt the reaction mixture was cooled to 0° C., quenched withaqueous 0.2N HCl (1.15 eq., 40.3 mL, 8.07 mmol) and diluted with water(40 mL) to produce a precipitation. The precipitation was filtered,washed with hexane-AcOEt (15 mL-2 mL) and dried under reduced pressureto afford pure colorless crystals of MK-046 (1.58 g, 62%).

Ph₃P (2.98 g, 11.37 mmol, 2.6 eq.) was dissolved in 30 mL of dry THF andcooled to 0° C. 40% DEAD in toluene (4.76 mL, 10.49 mmol, 2.4 eq.) wasadded and stirred for 30 min at 0° C. To the stirred solution wasdropwise added a mixture of MK-046 (1.58 g, 4.37 mmol) and2-(trimethylsilyl)ethanol (0.94 mL, 6.56 mL, 1.5 eq.) in 25 mL of THF at0° C. After 30 min, the reaction mixture was warmed up to it graduallyover 1 hr. The resulting mixture was evaporated and purified bychromatography on silica gel (hexane/AcOEt: 6/1 to 5/1) to afford oil ofMK-047 (2.01 g, 99%).

MK-047 (2.01 g, 4.35 mmol) was dissolved in 60 mL of AcOEt. Then 10%Pd/C (50% wet, 200 mg) was added. H₂ balloon was mounted and the mixturewas purged with H₂ (1 atm). After stirred overnight at rt it was workedup in usual manner and purified by chromatography on silica gel(hexane/AcOEt: 6/1 to 5/1) to afford colorless crystals of MK-048 (1.20g, 84%).

Using similar procedure for the synthesis of 343-YW-203 from(S)-1,3-butanediol, (R)-1,3-butanediol (9.80 g, 108.7 mmol) wasconverted to MK-049 (21.54 g, 94% 2 steps) as compound purified.

Using similar procedure for the synthesis of 343-YW-276 from 343-YW-203,MK-049 (15.57 g, 74.05 mmol) was converted to MK-050 (19.51 g, 72% 2steps) as compound purified.

To a stirred solution of MK-050 (5.15 g, 14.16 mmol) in 35 mL of dry THFwas added n-BuLi in hexane (1.6M, 19.5 mL, 31.14 mmol) at −78° C. After1 hr the reaction mixture was quenched with saturated aqueous solutionof NH₄Cl and diluted with AcOEt. The organic extract was washed withbrine, dried over MgSO₄, filtered and concentrated to give crudeproduct. It was purified by chromatography on silica gel (hexane/AcOEt:6/1 to 5/1) to afford oil of MK-051 (2.69 g, 93%).

Using similar procedure for the synthesis of NY-01 from TM-03, TM-03(2.79 g, assumed to contain 10.0 mmol) coupled with MK-051 (2.68 g, 13.1mmol) was converted to crude alcohol. The crude product was purified bychromatography on silica gel (hexane/AcOEt: 5/1 to 3/1) to afford oil ofMK-052 (less polar single isomer, 986 mg, 20%) and oil of MK-053 (polarsingle isomer, 1.48 g, 30%).

Using similar procedure for the synthesis of TM-05 from TM-04, MK-052(less polar single isomer, 968 mg, 1.96 mmol) was converted to colorlessoil of MK-054 and purified (single isomer, 870 mg, 90%).

Using similar procedure for the synthesis of MK-024 from MK-023, MK-054(834 mg, 1.69 mmol) treated with 3-MPMCl (0.61 mL, 4.21 mmol) wasconverted to colorless oil of MK-055 and purified (984 mg, 95%).

To a stirred solution of MK-055 (998 mg, 1.62 mmol) in 16 mL of THF wasadded TBAF in THF (1M, 2.43 mL, 2.43 mmol) at rt. After 3 hrs themixture was worked up in usual manner and purified by chromatography onsilica gel (hexane/AcOEt: 3/1) to afford colorless oil of MK-056 (696mg, 86%).

Using similar procedure for the synthesis of MK-008 from MK-007, MK-056(695 mg, 1.39 mmol) was converted to crude aldehyde of MK-057 (728 mg).The crude aldehyde was used for next step without purification.

Using similar procedure for the synthesis of MK-044 from MK-043, MK-057(728 mg, assumed to contain 1.39 mmol) was converted to colorless oil ofMK-058 and purified (643 mg, 90% 2 steps).

To a solution of MK-058 (200 mg, 0.389 mmol) and2,6-(^(t)Bu)₂-4-Me-pyridine (798 mg, 10 eq., 3.89 mmol) in 5 mL of dryCH₂Cl₂ was added (COCl)₂ in CH₂Cl₂ (2M, 0.97 ml, 5 eq., 1.94 mmol) at 0°C. and the solution was allowed to warm to rt. After stirred for 45 minthe reaction mixture was concentrated in vacuo under nitrogen atmosphereto give crude product including acid chloride MK-059. The crude productwas used for next step without purification.

The crude product including MK-059 (assumed to contain 0.389 mmolderived from 0.389 mmol of MK-058, 1.03 eq.) was dissolved in 4 mL ofdry CH₂Cl₂ at 0° C. A solution of MK-048 (124 mg, 0.377 mmol) in 4 mL oftoluene was added and the mixture was allowed to warm to it. Afterstirred for 15 min the reaction mixture was quenched with saturatedaqueous solution of NaHCO₃ and extracted with AcOEt. The organic extractwas washed with brine, dried over Na₂SO₄, filtered and concentrated togive crude product. It was purified by chromatography on silica gel(hexane/AcOEt: 3/1) to afford pale brown oil of MK-060 (297 mg, 96%).

To a stirred mixture of MK-060 (194 mg, 0.235 mmol) in 4 mL of CH₂Cl₂and 0.2 mL of aqueous phosphate buffer (pH 6.86) was added DDQ (59 mg,1.1 eq., 0.259 mmol) at 0° C. After stirred for 1.5 hrs at 0° C. thereaction mixture was quenched with aqueous solution of NaHCO₃ andextracted with AcOEt. The organic extract was washed with saturatedaqueous solution of NaHCO₃ and brine, dried over Na₂SO₄, filtered andconcentrated to give crude oil. The crude product was purified bychromatography on silica gel (hexane/AcOEt: 5/2) to afford colorless oilof MK-061 (138 mg, 83%).

Using similar procedure for the synthesis of MK-031 from MK-030, MK-061(165 mg, 0.234 mmol) was converted to crude MK-062 (159 mg, >100%). Thecrude MK-062 was used for next step without purification.

Ph₃P (221 mg, 0.844 mmol, 3.6 eq.) was dissolved in 39 mL of dry THF andcooled to 0° C. 40% DEAD in toluene (0.32 mL, 0.703 mmol, 3.0 eq.) wasadded and stirred for 20 min at 0° C. To the stirred solution was addeddropwise over 15 min a solution of crude MK-062 (159 mg, assumed tocontain 0.234 mmol) in 39 mL of THF at 0° C. After 10 min at 0° C. thereaction mixture was evaporated and purified by chromatography on silicagel (hexane/AcOEt: 2/1 to 3/2) to afford oil of MK-063 (115 mg, 84% 2steps).

To a stirred mixture of MK-063 (115 mg, 0.196 mmol) in 5 mL of CH₂Cl₂and 0.5 mL of aqueous phosphate buffer (pH 6.86) was added DDQ (103 mg,2.3 eq., 0.452 mmol) at 0° C. and the mixture was allowed to warm to rt.After stirred for 24 hrs at rt the reaction mixture was worked up in theusual manner to give a crude oil. The crude product was purified bychromatography on silica gel (hexane/AcOEt: 2/1 to 1/3) to affordcolorless oil of MK-064 (2^(nd) elution, 14 mg, 15%), colorless oil ofMK-065 (3^(rd) elution, 21 mg, 23%), and colorless oil of MK-066 (1^(st)elution, 14 mg, 15%).

Using similar procedure for the synthesis of NF0552 from MK-021, MK-065(21 mg, 0.0451 mmol) treated at low temperature was converted tocolorless oil of MK-066 (15 mg, 72%) as compound purified.

Using similar procedure for the synthesis of NF1226 from MK-035, MK-066(29 mg, 0.0626 mmol) was converted to crude pale yellow crystals. Thecrude product was purified by chromatography on silica gel(hexane/AcOEt=1/3 to AcOEt alone) to afford colorless crystals of NF1535(17.6 mg, 74%).

Using similar procedure for the synthesis of NF1226 from MK-035, MK-064(14 mg, 0.0302 mmol) was converted to crude pale yellow crystals. Thecrude product was purified by chromatography on silica gel(CH₂Cl₂/AcOEt=4/1, 2/1, to 1/1) to afford colorless crystals of NF1537(8.2 mg, 72%).

Synthetic Procedure for NF2306

To a stirred solution of methyl (R)-(−)-3-hydroxy-2-methylpropionate(7.00 g, 59.26 mmol) in 66 mL of CH₂Cl₂ and 132 mL of cyclohexane wereadded CCl₃C(═NH)OBn (13.2 mL, 71.1 mmol) and CF₃SO₃H (cat. 0.2 mL) atrt. After 3 hrs the reaction mixture was diluted with hexane to formprecipitation. After filtering precipitation off, filtrates was washedwith saturated aqueous NaHCO₃ and brine, dried over Na₂SO₄, filtered,and concentrated to give a crude oil. The crude product was purified bychromatography on silica gel (hexane/AcOEt=20/1) to afford colorless oilof MK-067 (9.60 g, 78%).

LiAlH₄ (2.62 g, 69.1 mmol) was suspended in 250 mL of dry THF. To thesuspension was dropwise added a solution of MK-067 (9.59 g, 46.0 mmol)in 57 mL of dry THF at 0° C. After stirred for 1 hr at 0° C. thereaction mixture was quenched with MeOH (13 mL), water (2.5 mL), 10%NaOH (2.5 mL), then water (7.5 mL). The mixture was dried over MgSO₄,filtered and evaporated to a crude product. The crude product waspurified by chromatography on silica gel (hexane/AcOEt=2/1) to affordcolorless oil of MK-068 (7.89 g, 95%).

Using similar procedure for the synthesis of MK-008 from MK-007, MK-068(7.89 g, 43.76 mmol) was converted to crude MK-069 (8.66 g, >100%). Thecrude MK-069 was used for next step without purification.

To a stirred suspension of CuI (10.83 g, 56.9 mmol, 1.3 eq.) in 100 mLof dry Et₂O was added over 15 min MeLi in Et₂O (1.14 M, 98.6 mL, 112.5mmol, 2.57 eq.) at 0° C. After stirred for 30 min at 0° C., the mixturewas cooled to −78° C. Crude MK-069 (8.66 g, assumed to contain 43.76mmol) in 75 mL of dry Et₂O was added over 40 min at −78° C. then themixture was stirred at −78° C. for additional 1 hr. The reaction mixturewas allowed to warm to −20° C. over 1.5 hr, then quenched with 28%aqueous NH₃ solution and extracted with AcOEt. The organic extract waswashed with brine, dried over MgSO₄, filtered and concentrated to give acrude oil. The crude product was purified by chromatography on silicagel (hexane/AcOEt: 5/1) to afford pale yellow oil of MK-070 (6.89 g, 81%2 steps).

Ph₃P (9.24 g, 35.24 mmol, 1.4 eq.) was dissolved in 70 mL of dry THF andcooled to 0° C. 40% DEAD in toluene (14.84 mL, 32.72 mmol, 1.3 eq.) wasadded and stirred for 20 min at 0° C. To the stirred solution wasdropwise added a solution of MK-070 (4.89 g, 25.17 mmol) and PhCO₂H(4.00 g, 32.7 mmol, 1.3 eq.) in 30 mL of dry THF at 0° C. After 30 minat 0° C. the mixture was allowed to warm to it overnight. The resultingreaction mixture was evaporated and diluted with hexane-AcOEt. Afterfiltration of generated precipitate, the filtrate was concentrated toyield crude oil. The crude product was purified by chromatography onsilica gel (hexane/AcOEt: 15/1) to afford pale yellow oil of MK-071(6.84 g, 91%).

To a stirred solution of MK-071 (6.84 g, 22.91 mmol) in 38 mL of EtOHwas added aqueous 3N—NaOH (15.3 mL, 45.81 mmol) then the mixture wasstirred at 80° C. for 1 hr. The resulting mixture was evaporated,extracted with Et₂O, washed with brine, dried over MgSO₄, filtered andconcentrated to yield a crude oil. The crude product was purified bychromatography on silica gel (hexane/AcOEt: 5/1 to 5/3) to affordcolorless oil of MK-072 (4.17 g, 94%).

To a suspension of 66% NaH (916 mg, 25.20 mmol) in 30 mL of DMF wasadded a solution of MK-072 (2.72 g, 14.00 mmol) in 10 mL of DMF at 0° C.After stirring at 0° C. for 30 min 4-MPMCl (3.80 mL, 28.00 mmol) wasadded, then the mixture was allowed to warm to rt. After 2 days thereaction was quenched with saturated aqueous solution of NH₄Cl andextracted with AcOEt. The organic extract was washed with saturatedaqueous solution of NaHCO₃ and brine, dried over Na₂SO₄, filtered andconcentrated. The crude product was purified by chromatography on silicagel (Hexane/AcOEt: 12/1) to afford colorless oil of MK-073 (3.88 g,88%).

50 wt % suspension of Raney-Ni (W2) in basic water (9.5 g) was addedinto a flask, then the suspension was washed with water and EtOH. Tothis suspension was added a solution of MK-073 (3.88 g, 12.34 mmol) in150 mL of EtOH. H₂ ballon was mounted and the mixture was purged withH₂. After stirred for 5 days under H₂ (1 atm) at rt, the reactionmixture was filtered through Celite and the filtrate was evaporated togive a crude oil. The crude product was purified by chromatography onsilica gel (Hexane/AcOEt: 3/1) to afford colorless oil of MK-074 (2.61g, 94%).

Using similar procedure for the synthesis of MK-008 from MK-007, MK-074(2.61 g, 11.63 mmol) was converted to crude oil of MK-075 (2.70 g,quant.). The crude MK-075 was used for next step without purification.

Using similar procedure for the synthesis of 343-YW-276 from 343-YW-203,crude MK-075 (2.70 g, assumed to contain 11.63 mmol) was converted toMK-076 (3.92 g, 89% 2 steps).

MK-076 (2.03 g, 5.37 mmol, 1.36 eq.) was dissolved in 27 mL of dry THFand cooled to −78° C. under nitrogen. n-BuLi in hexane (1.6M, 6.71 mL,10.73 mmol, 2.71 eq.) was added and stirred at −78° C. for 1 hr. Asolution of crude TM-03 (1.11 g, assumed to contain 3.96 mmol) in 7 mLof dry THF was dropwise added to the mixture and stirred for 30 min at−78° C. The reaction mixture was allowed to warm to 10° C. slowly over2.5 hrs. The mixture was quenched with saturated aqueous solution ofNH₄Cl and extracted with AcOEt. The organic extract was washed withsaturated aqueous solution of NaHCO₃ and brine, dried over Na₂SO₄,filtered and concentrated. The crude product was purified bychromatography on silica gel (hexane/AcOEt: 7/1 to 4/1) to afford oil ofMK-077 (less polar single isomer, 541 mg, 27%) and oil of MK-078 (polarsingle isomer, 1.21 g, 60%).

Using similar procedure for the synthesis of TM-05 from TM-04, MK-077(less polar single isomer, 3.85 g, 7.61 mmol) was converted to colorlessoil of MK-079 (single isomer, 3.38 g, 87%).

Using similar procedure for the synthesis of MK-024 from MK-023, MK-079(3.38 g, 6.63 mmol) treated with 3-MPMCl (2.89 mL, 19.9 mmol) wasconverted to colorless oil of MK-080 (3.86 g, 92%).

Using similar procedure for the synthesis of MK-056 from MK-055, MK-080(3.85 g, 6.12 mmol) was converted to colorless oil of MK-081 (3.00 g,95%).

Using similar procedure for the synthesis of MK-058 from MK-056, MK-081(1.82 g, 3.54 mmol) was converted to colorless oil of MK-083 (1.80 g,96% 2 steps).

Using similar procedure for the synthesis of MK-059 from MK-058, MK-083(1.80 g, 3.40 mmol) was converted to crude oil of MK-084. The crudeMK-084 was used for next step without purification.

Using similar procedure for the synthesis of MK-060 from MK-059, crudeMK-084 coupled with MK-048 (853 mg, 2.60 mmol) was converted to palebrown oil of MK-085 (2.14 g, 98% 2 steps).

Using similar procedure for the synthesis of MK-061 from MK-060, MK-085(2.14 g, 2.55 mmol) was converted to pale yellow oil of MK-086 (1.74 g,95%).

Using similar procedure for the synthesis of MK-031 from MK-030, MK-086(1.74 g, 2.42 mmol) was converted to crude oil of MK-087 (1.57 g,quant.). The crude MK-087 was used for next step without purification.

Using similar procedure for the synthesis of MK-063 from MK-062, crudeMK-087 (1.57 g, assumed to contain 2.42 mmol) was converted to paleyellow oil of MK-088 (1.68 g, including ca 0.38 g of inseparableimpurity derived from DEAD, ca 90% 2 steps).

Using similar procedure for the synthesis of MK-065 from MK-063, MK-088(1.68 g, including ca 0.38 g of inseparable impurity derived from DEAD,assumed to contain 2.17 mmol) was converted to colorless solid of MK-089(525 mg, 50%).

Using similar procedure for the synthesis of NF0552 from MK-021, MK-089(458 mg, 0.955 mmol) was treated with Dess-Martin reagent at lowtemperature to give a pale yellow solid of MK-090 (250 mg, 55%).

MK-090 (250 mg, 0.524 mmol) was dissolved in 3.5 mL of CH₂Cl₂ and cooledto 0° C. A mixture of 50% Hydrofluoric acid (24N, 3.5 mL) and 14 mL ofCH₃CN was added to the solution and stirred for 1 hr at 0° C. before themixture was allowed to warm to 15° C. slowly over 1.5 hrs. Then, thereaction mixture was poured into a stirred biphasic solution ofsaturated aqueous solution of NaHCO₃ and AcOEt. The organic extract waswashed with brine, dried over Na₂SO₄, filtered and concentrated to crudeproduct. The crude product was purified by chromatography on silica gel(CH₂C₂/MeOH: 13/1) to afford colorless crystals of NF2306 (174 mg, 42%).

Preparation of C13-Oxygen and Fluoro Analog, NF2432, NF2544, NF2547,NF2553 and NF2556

Synthetic Procedure for NF2432

1) Preparation of the Macrocyclic Part

To a stirred THF solution of dibromide YE-43 (18.6 g, 37 mmol), whichwas prepared using similar procedure for the intermediate 554-RB-228from methyl (S)-3-hydroxybutyrate (47%, 5 steps), was addedn-butyllithium (1.6M in hexane solution, 47 ml, 75 mmol) at −78° C. Themixture was allowed to warm to 0° C. for 30 min and then stirred at −78°C. for additional 30 min. A THF solution of aldehyde NY-20 (6.17 g, 24mmol) was added, the mixture was allowed to warm to 0° C. and stirred at0° C. for 30 min, after which a saturated solution of NH₄Cl was added.The mixture was extracted with EtOAc and the organic extract was washedwith a saturated solution of NaHCO₃, brine, dried over anhydrous Na₂SO₄,filtered and concentrated. The crude product was purified bychromatography on silica gel using 12% EtOAc/hexane to give 3.55 g (5.9mmol, 25%) of the less polar isomer YE-01 and 5.5 g (0.093 mol, 39%) ofthe more polar isomer YE-02.

To a solution of YE-02 (10.1 g, 17.0 mmol) in n-hexane (170 mL),quinoline (0.25 eq., 4.25 mmol, 0.50 mL) and 5 wt. % Pd on BaSO₄ (0.05eq., 0.85 mmol, 1.81 g) was added at rt. And the reaction mixture waspurged with H₂ and stirred under H₂ atmosphere for 11 hrs. The catalystwas filtered off and the filtrate was concentrated. The crude product(11.5 g) was purified by chromatography on silica gel using 16%EtOAc/hexane to give 8.19 g (13.7 mmol, 81% 2 steps) of the desiredallylic alcohol YE-03.

The alcohol YE-03 (8.04 g, 13.5 mmol) was dissolved in CH₂Cl₂ (200 mL),2,6-lutidine (7.8 mL, 67.0 mmol) was added and the mixture was cooled to0° C. in ice/water bath. Then TBSOTf (7.7 mL, 33.5 mmol) was added andthe mixture was allowed to warm to rt. After 2 hrs, it was cooled to 0°C. and was quenched with MeOH and a saturated solution of NaHCO₃. Themixture was extracted with EtOAc, the organic layer was washed with asaturated solution of NaHCO₃, 5% citric acid aq., a saturated solutionof NaHCO₃, and brine. The organic layer was dried over Na₂SO₄, filteredand concentrated. The crude product was purified by chromatography onsilica gel using 2-4% EtOAc/hexane to give 9.60 g (13.5 mmol, quant.) ofYE-04.

To a stirred ether solution (200 ml) of YE-04 (9.60 g, 13.5 mmol) at 0°C. was added lithium tetrahydroborate (0.60 g, 27.5 mmol). The mixturewas allowed to warm to rt and stirred for 2 days. Then the mixture wascooled to 0° C. and lithium tetrahydroborate (0.30 g, 13.8 mmol) wasadded again. The mixture was allowed to warm to rt and stirredovernight. The mixture was cooled to 0° C., then a saturated solution ofNH₄Cl (2 ml) was added slowly. After the stirring for 20 min, asaturated solution of NH₄Cl (100 ml) was added. The mixture wasextracted with EtOAc and the organic extract was washed with a saturatedsolution of NH₄Cl, brine, dried over anhydrous Na₂SO₄, filtered andconcentrated. The crude product was purified by chromatography on silicagel using 12-15% EtOAc/hexane to give 8.51 g (13.5 mmol, quant.) of thealcohol YE-05.

To a stirred solution of YE-05 (1.90 g, 3.03 mmol) in toluene (45 ml)was added triphenylphosphine (1.6 g, 6.1 mmol), a mixture of diethylazodicarboxylate (40% in toluene, 2.1 ml, 4.6 mmol) and iodomethane(0.29 ml, 4.7 mmol). The mixture was stirred for 40 min after which amixture of diethyl azodicarboxylate (in toluene, 0.5 ml, 1.1 mmol) andiodomethane (0.06 ml, 1.2 mmol) was added. The mixture was stirred for20 min and the solvent was evaporated in vacuo. The concentrate waspurified by chromatography on silica gel using 1-1.5% EtOAc/hexane togive 2.05 g (2.78 mmol, 92%) of the iodide YE-06.

2) Preparation of the Aromatic Part

3-Bromo-4-hydroxy-5-methoxybenzaldehyde (24.8 g, 0.107 mol) wasdissolved in DMF (400 ml). K₂CO₃ (20 g, 0.14 mol) and iodomethane (8.8ml, 0.14 mol) was added. The mixture was stirred for 4 hrs and was thencooled to 0° C. in ice/water bath and diluted with ether (300 ml). Thenice-water (600 ml) was added slowly. The mixture was extracted withether and the organic extract was washed with water, brine, dried overanhydrous Na₂SO₄, filtered and concentrated to give 22.1 g (90 mmol,84%.) of YE-07.

YE-07 (13.3 g, 54.2 mmol) was dissolved in chloroform (350 mL) undernitrogen. m-CPBA (>70%, 31 g, 126 mmol) was added and the solution wasgently refluxed for 1 hr. The reaction mixture was cooled to 0° C. andwas poured into saturated stirred solution of NaHCO₃. After stirring for15 min, the organic layer was separated, washed with saturated Na₂SO₃,saturated NaHCO₃, dried over Na₂SO₄, filtered and concentrated

The concentrate was dissolved in MeOH (150 ml) and 6N HCl (150 ml) wasadded. The mixture was stirred for 15 min. and partially concentrated.The mixture was extracted with EtOAc and the organic extract was washedwith water, dried over Na₂SO₄, filtered and concentrated. to give 10.8 gof crude phenol.

The crude phenol was methylated using the same procedure as thepreparation of YE-07 to give 9.8 g (39.7 mmol, 73%) of YE-08.

To a stirred ether solution of bromobenzene YE-08 (14.6 g, 59.1 mmol) at−78° C. was added n-butyllithium (1.6M in hexane solution, 50 ml, 1.3eq., 80 mmol), and the mixture was stirred at −78° C. for 1.5 hrs. Asolution of iodomethane (10 ml, 161 mmol) in ether (20 ml) was added,and the mixture was allowed to warm to rt and stirred for 1.5 hrs, afterwhich a saturated solution of NH₄Cl was added. The mixture was extractedwith ether and the organic extract was washed with brine, dried overanhydrous Na₂SO₄, filtered and concentrated. The concentrate waspurified by chromatography on silica gel using 6-9% EtOAc/hexane to give9.6 g (52.6 mmol, 89%) of 2,3,5-trimethoxytoluene.

The toluene 9.6 g (52.6 mmol, 89%) was dissolved in DME (130 mL) andcopper (II) bromide (25 g, 112 mmol) was added portionwise over 6 hrs.After stirring another 1 hr, the reaction mixture was filtrated and thefiltrate was concentrated. The concentrate was purified bychromatography on silica gel using 5-12% EtOAc/hexane to give 12.7 g(48.6 mmol, 92%) of YE-09.

To a stirred ether solution of YE-09 (45.6 mmol, 11.9 g) at −78° C. wasadded n-butyllithium (1.6M in hexane solution, 37 ml, 59 mmol) and themixture was stirred at −78° C. for 1 hr. A finely crushed dry-ice wasadded slowly and the mixture was allowed to warm to −10° C. After 2 hrs,the reaction was quenched with water (200 ml). The mixture was washedwith ether then acidified with 1N HCl. The mixture was extracted withEtOAc and the organic extract was washed with water, brine, dried overanhydrous Na₂SO₄, filtered and concentrated to give 9.7 g (42.8 mmol,94%) of YE-10.

To a stirred solution of YE-10 (1.4 g, 6.3 mmol) in dry CH₂Cl₂ (40 mL)was added BBr₃ (1M solution, 28 mL, 28 mmol) at −78° C. under nitrogenatmosphere and the mixture was allowed to warm to rt. After 8 hrs, themixture was cooled to 0° C. and poured into water. The organic layer wasseparated and washed with a solution of 5% glycerol, brine, dried overanhydrous Na₂SO₄, filtered and concentrated to give 0.9 g of the crudeproduct.

To a stirred solution of the crude product (0.9 g, 4.8 mmol) in dryacetonitrile (20 mL) were added MeOH (0.78 ml) followed byN,N-diisopropylethylamine (1.7 ml, 9.7 mmol). To the mixture was addedTMSCHN₂ (2.0M in hexane, 4.8 mL, 9.6 mmol) and the mixture was stirredat 30° C. After 1 hr., the mixture was cooled to 0° C., poured intowater and extracted with EtOAc. The organic extract was washed with asaturated solution of NH₄Cl, water, brine, dried with anhydrous Na₂SO₄,filtered and concentrated. The crude product was purified bychromatography on silica gel using 6% EtOAc/hexane to give 0.37 g (1.74mmol, 28% 2 steps) of YE-11.

To a stirred suspension of NaH (0.16 g, 4.4 mmol) in dry THF (10 mL) wasadded YE-11 (0.37 g, 1.7 mmol) in dry THF (8 mL) at 0° C. After stirringfor 30 min at 0° C., TBDPSCl (0.5 ml, 1.9 mmol) was added. The mixturewas allowed to warm to rt and stirred for 15 min. The mixture was cooledto 0° C., poured into water and extracted with EtOAc. The organicextract was washed with water, brine, dried with anhydrous Na₂SO₄,filtered and concentrated. The concentrate was purified bychromatography on silica gel using 6% EtOAc/hexane to give 0.63 g (1.39mmol, 80%) of silyl ether.

To a stirred solution of the silyl ether (3.7 g, 8.3 mmol) in DMF (50mL) was added Cs₂CO₃ (3.0 g, 9.2 mmol) and iodomethane (1.3 mL, 20.8mmol). The mixture was stirred overnight and was cooled to 0° C. inice/water bath. Then the mixture was poured into ice-cold, saturatedsolution of NH₄Cl (100 mL) and extracted with EtOAc. The organic extractwas washed with water, brine, dried over anhydrous Na₂SO₄, filtered andconcentrated. The concentrate was purified by chromatography on silicagel using 5% EtOAc/hexane to give 2.83 g (6.09 mmol, 73%) of YE-12.

Using similar procedure for the intermediate 10 of NF2561, YE-12 (0.30g, 0.64 mmol) was converted to YE-13 (0.29 g, 0.51 mmol, 80% 2 steps).

YE-13 (0.29 g, 0.51 mmol) was dissolved in THF (10 mL). Then 1.0Msolution of TBAF in THF (1.5 mL, 1.5 mmol) were added at rt. The mixturewas stirred overnight after which a saturated solution of NH₄Cl wasadded. The mixture was extracted with EtOAc and the organic extract waswashed with a saturated solution of NaHCO₃, water, brine, dried withanhydrous Na₂SO₄, filtered and concentrated. The crude product waspurified by chromatography on silica gel using 10% EtOAc/hexane to EtOAcas eluents to give 0.17 g (0.51 mmol, quant.) of phenol.

To a stirred suspension of sodium hydride (0.28 g, 7.7 mmol) in dry THF(10 mL) was added the phenol (2.0 g, 5.9 mmol) in dry THF (20 mL) at 0°C. After stirring for 15 min, chloromethyl methyl ether (0.57 mL, 7.5mmol) at 0° C. After 3 hr, the mixture was poured into a saturatedsolution of NH₄Cl and extracted with EtOAc. The organic extract waswashed with a saturated solution of NaHCO₃, brine, dried over anhydrousNa₂SO₄, filtered and concentrated. The crude product was purified bychromatography on silica gel using 15% EtOAc/hexane to give 1.4 g (3.7mmol, 62%) of YE-14.

Using similar procedure for the intermediate 14 of NF2561, YE-14 (1.4 g,3.7 mmol) was converted to YE-15 (1.6 g, 3.4 mmol, 93% 2 steps).

Using similar procedure for the intermediate 18 of NF2561, the iodide(502 mg, 0.68 mmol) was converted to YE-16 (315 mg, 0.33 mmol, 48% 3steps).

YE-16 (310 mg, 0.32 mmol) was dissolved in THF (12 mL). Then, 1.0Msolution of tetrabutylammonium fluoride in THF (1.6 mL, 1.6 mmol) wasadded at 0° C. The mixture was stirred at rt for 2 days after which 10%KHSO₄ solution was added. The mixture was extracted with EtOAc and theorganic extract was washed with water, brine, dried with anhydrousNa₂SO₄, filtered and concentrated. The crude product was dried byazeotropic distillation with toluene to give 270 mg of YE-17 with silylimpurity.

To a stirred solution of crude YE-17 (270 mg) in THF (20 mL) were addedtriethylamine (0.090 mL, 0.64 mmol) and 2,4,6-trichlorobenzoyl chloride(0.085 mL, 0.54 mmol) at rt. After 16 hrs, the reaction mixture wasdiluted with toluene (300 mL) and added dropwise to a solution of4-(dimethylamino)pyridine (980 mg, 8.0 mmol) in toluene (320 mL) over aperiod of 6 hrs under reflux. The resultant mixture was stirred for 0.5hr under reflux. After concentration under reduced pressure, the residuewas dissolved in EtOAc and washed with 10% KHSO₄ aq sol., water, brineand dried over anhydrous Na₂SO₄, filtered and concentrated. The crudeproduct was purified by chromatography on silica gel using 6-30%EtOAc/hexane to give 114 mg (0.23 mmol, 72% 3 steps) of YE-18.

Using similar procedure for 509-HD-125, YE-18 (20 mg, 0.041 mmol) wasconverted to YE-19 (22 mg, quant.).

To a stirred solution of YE-19 (22 mg, 0.041 mmol) in THF (1.4 mL)-H₂O(0.7 mL) was added trifluoroacetic acid (1.4 mL) at 0° C. The mixturewas then allowed to warm to rt. After 1.5 hrs, the mixture was pouredinto a saturated solution of NaHCO₃ and extracted with EtOAc. Theorganic extract was washed with water, brine and dried over anhydrousNa₂SO₄, filtered and concentrated. The crude product was purified bychromatography on silica gel using 30% EtOAc/hexane to give 13.3 mg(0.033 mmol, 81%) of NF2432.

Synthetic Procedure for NF2544

To a stirred suspension of NaH (2.3 g, 63 mmol) in dry THF (100 mL) wereadded 2-bromo-4-fluorophenol (10 g, 52 mmol) in thy THF (20 mL) at 0° C.After stirring for 30 min at 0° C., chloromethyl methyl ether (4.8 mL,63 mmol) was added. The mixture was allowed to warm to rt and stirredfor 1.5 hrs. The mixture was cooled to 0° C., poured into water andextracted with EtOAc. The organic extract was washed with water, brine,dried with anhydrous Na₂SO₄, filtered and concentrated. The concentratewas purified by chromatography on silica gel using 10% EtOAc/hexane togive 10.7 g (45.5 mmol, 87%) of YE-20.

To a stirred suspension of YE-20 (10.7 g, 46 mmol) in dry ether (150 mL)was added 1.6M n-BuLi in hexane (34 mL, 54.4 mmol) at −78° C. undernitrogen atmosphere. After 1 hr, dry DMF (15 mL) was added and themixture was allowed to warm to it. It was quenched with a saturatedsolution of NH₄Cl and extracted with EtOAc. The organic extract waswashed with a saturated solution of NH₄Cl, water, brine, dried overanhydrous Na₂SO₄, filtered and concentrated. The crude product waspurified by chromatography on silica gel using 10% EtOAc/hexane to give7.8 g (42 mmol, 93%) of YE-21.

To a stirred solution of YE-21 (6.8 g, 37 mmol) in toluene (200 mL) wasadded ethylene glycol (12 g, 193 mmol) and p-toluenesulfonic acidmonohydrate (0.3 g) at rt. and refluxed using Dean-Stark apparatus.After 4 hr, the mixture was cooled to 0° C. in ice/water bath. Then TEA(15 ml, 0.10 mol) was added, the mixture was stirred for 10 min, thenpoured into a saturated solution of NaHCO₃. The mixture was extractedwith EtOAc and the organic extract was washed with a saturated solutionof NaHCO₃, brine, dried over anhydrous Na₂SO₄, filtered and concentratedto give 6.7 g of phenol as crude product.

To a stirred suspension of NaH (1.5 g, 41 mmol) in dry THF (120 mL) wereadded the phenol (6.7 g, 36 mmol) in dry THF (20 mL) at 0° C. Afterstirring for 10 mm at 0° C., TBSCl (6.3 g, 42 mmol) was added. Themixture was allowed to warm to rt and stirred overnight. The mixture wascooled to 0° C., poured into water and extracted with EtOAc. The organicextract was washed with water, brine, dried over anhydrous Na₂SO₄,filtered and concentrated. The concentrate was purified bychromatography on silica gel using 2% EtOAc/hexane (containing 0.2% oftriethylamine) to give 8.75 g (29.3 mmol, 79% 2 steps) of YE-22.

To a stirred suspension of YE-22 (20 g, 67 mmol) in THF (350 mL) wasadded 1.6M n-BuLi in hexane (50 mL, 80 mmol) at −78° C. under nitrogenatmosphere. After 1 hr TMEDA (15 mL, 99 mmol) was added and stirred at−78° C. for another 10 min. To the mixture was added dry DMF (5.0 mL, 65mmol) and the mixture was allowed to warm to −20° C. and stirred for 40min. It was quenched with AcOH (14 mL), and poured into water. Themixture was extracted with EtOAc and the organic extract was washed witha saturated solution of NaHCO₃, brine, dried over anhydrous Na₂SO₄,filtered and concentrated. The crude product was purified bychromatography on silica gel using 2-5% EtOAc/hexane to give 15.8 g(48.4 mmol, 72%) of YE-23.

To a stirred MeOH solution (350 mL) of YE-23 (15.8 g, 48.4 mmol) at 0°C. was added NaBH₄ (2.0 g, 53 mmol). The mixture was stirred at 0° C.for 30 min. Then the mixture was quenched with AcOH (10 mL), and pouredinto a saturated solution of NaHCO₃. The mixture was extracted withether and the organic extract was washed with a saturated solution ofNaHCO₃, brine, dried over anhydrous Na₂SO₄, filtered and concentrated togive 16.4 g (48.4 mmol, quant.) of alcohol.

To a stirred acetone solution (300 mL) of the alcohol (16.4 g, 48.4mmol) at 0° C. was added 1N HCl (8 mL, 8 mmol). The mixture was stirredat 0° C. for 70 min, then was poured into a saturated solution ofNaHCO₃. The mixture was partially concentrated to remove acetone and theconcentrate was extracted with ether. The organic extract was washedwith brine, dried over anhydrous Na₂SO₄, filtered and concentrated togive 13.6 g (47.8 mmol) of YE-24.

YE-24 (13.6 g, 47.8 mmol) was dissolved in 900 mL of dichloromethane andPDC (53.7 g, 143 mmol) were added to the solution. The reaction mixturewas stirred for 7 d at rt and the mixture was diluted with ether (300mL). After passing through Florisil column, YE-25 was obtained ascolorless oil (11.7 g, 86%, 3 steps).

YE-25 (11.7 g, 41.4 mmol) was dissolved in THF (120 mL), 1.7Nhydrochloric acid (60 mL, 100 mmol) were added and the mixture wasstirred at 50° C. for 1 hrs, then refluxed for 3 hrs. The mixture wascooled to rt and filtrated. To the filtrated was added 5N NaOH (18 mL)and the organic layer was separated. The aqueous layer was saturatedwith NaCl and was extracted with EtOAc. The combined organic extract wasconcentrated and the concentrate was triturated with ether to givephenol (4.9 g, 32.2 mmol).

Using similar procedure for TM-37, the phenol (4.2 g, 27.6 mmol) wasconverted to YE-26 (4.2 g, 21.2 mmol, 77%).

Using similar procedure for TM-50, YE-26 (4.1 g, 20.9 mmol) wasconverted to YE-27 (4.9 g, 14.6 mmol, 70% 3 steps).

Using similar procedure for TM-39, YE-27 (1.5 g, 4.5 mmol) was convertedto YE-28 (1.60 g, 3.8 mmol, 85% 2 steps).

Using similar procedure for the intermediate 18 of NF2561, the iodide(510 mg, 0.69 mmol) was converted to YE-29 (690 mg, 0.67 mmol, 97% 3steps).

Using similar procedure for the intermediate YE-17, YE-29 (520 mg, 0.57mmol) was converted to YE-30 (430 mg) with silyl impurity.

Using similar procedure for YE-18, YE-30 (430 mg) was converted to YE-31(94 mg, 0.21 mmol, 36.5% 3 steps).

Using similar procedure for ER803064, YE-31 (45 mg, 0.099 mmol) wasconverted to NF-2544 (13 mg, 0.036 mmol, 36% 2 steps).

Synthetic Procedure for NF2547

5-Bromovanillin (20.0 g, 86.56 mmol) and AlCl₃ (12.70 g, 95.22 mmol)were dissolved in 145 mL of dry CH₂Cl₂. To the stirred solution wasdropwise added pyridine (30.80 mL, 380.9 mmol) over 10 min at rt(exothermic reaction). Then the mixture was warmed to 45° C. and stirredfor 20 hrs after which it was cooled to rt. The resulting mixture wasacidified with 3N HCl aq and extracted with AcOEt. The organic extractwas washed with brine, dried over Na₂SO₄, filtered and concentrated togive crude colorless crystals of MK-101 (18.4 g, <98%). The crude MK-101was used for next step without purification.

Crude MK-101 (13.15 g, assumed to contain 60.59 mmol), Br—CH₂—Cl (6.50mL, 96.9 mmol), and Cs₂CO₃ (31.59 g, 96.9 mmol) were suspended in 200 mLof DMF and the mixture was stirred at 110° C. for 20 hrs. The resultingmixture was diluted with water and extracted with AcOEt. The organicextract was washed with brine, dried over Na₂SO₄, filtered andconcentrated to give crude brown solid of MK-102. The crude product waspurified by chromatography on silica gel (hexane/AcOEt: 6/1 to 5/1) toafford colorless crystals of MK-102 (11.1 g, 77% 2 steps).

70% mCPBA (13.1 g, 53.1 mmol) was added to a stirred solution of MK-102(5.07 g, 22.12 mmol) in 110 mL of CHCl₃ and the mixture was heated up toreflux. After 3 hrs at reflux the mixture was cooled to 0° C., quenchedwith aqueous solution of Na₂SO₃, and extracted with CH₂Cl₂. The organicextract was washed with saturated aqueous solution of NaHCO₃ and brine,then evaporated to give crude pale brown solid of intermediate formate.

The crude formate was dissolved in 100 mL of MeOH. NaHCO₃ (3.7 g, 44mmol) was added and the suspension was stirred at rt for 30 min. Waterand AcOEt were added to the mixture and organic extract was washed withbrine, dried over Na₂SO₄, filtered and concentrated to give a crude oilysolid of MK-103. The crude product was purified by chromatography onsilica gel (hexane/AcOEt: 4/1) to afford colorless crystals of MK-103(3.78 g, 79% 2 steps).

Using similar procedure for the synthesis of 509-HD-209 from 509-HD-207,MK-103 (3.78 g, 17.40 mmol) was converted to colorless oil of MK-104(4.42 g, 97%).

To a stirred solution of MK-104 (2.00 g, 7.66 mmol) in 26 mL of dry Et₂Owas dropwise added n-BuLi in hexane (1.6M, 5.74 mL, 9.19 mmol, 1.2 eq.)at −78° C. After 1 hr dry DMF (1.20 mL, 15.3 mmol, 2.0 eq.) was added inone portion to the mixture and stirred at −78° C. for 30 min after whichsaturated aqueous solution of NH₄Cl was added. The resulting mixture wasextracted with AcOEt and the organic extract was washed with brine,dried over Na₂SO₄, filtered, and concentrated to give crude yellow oilof MK-105 (1.60 g). The crude product was used for next step withoutpurification.

The crude MK-105 (1.60 g, assumed to contain 7.66 mmol) was dissolved in26 mL of MeOH and the stirred solution was cooled to 0° C. NaBH₄ (290mg, 7.66 mmol) was added in some portions. After 20 min at 0° C. thereaction mixture was quenched with aqueous solution of NH₄Cl andextracted with AcOEt. The organic extract was washed with brine, driedover Na₂SO₄, filtered and concentrated to give crude oil. The crudeproduct was purified by chromatography on silica gel (hexane/AcOEt: 3/1to 2/1) to afford colorless crystals of MK-106 (1.44 g, 89% 2 steps).

To a stirred solution of MK-106 (1.26 g, 5.92 mmol) in 30 mL of DMF wasadded NBS (1.16 g, 6.51 mmol, 1.1 eq.) at 0° C. The mixture was stirredat 0° C. for 3 hrs before aqueous solution of Na₂SO₃ and AcOEt wereadded. The organic extract was washed with brine, dried over Na₂SO₄,filtered, and concentrated to give crude colorless crystals of MK-107(1.73 g, quant.). The crude product was used for next step withoutpurification.

Crude MK-107 (1.73 g, assumed to contain 6.51 mmol) and imidazole (1.01g, 14.80 mmol) were dissolved in 39 mL of DMF. TBS-Cl (1.78 g, 11.84mmol) was added to the solution and the mixture was stirred at rtovernight after which AcOEt was added. The resulting mixture was washedwith aqueous solution of NaHCO₃ and brine, dried over Na₂SO₄, filteredand concentrated to give crude crystals. The crude product was purifiedby chromatography on silica gel (hexane/AcOEt: 9/1) to afford colorlesscrystals of MK-108 (2.25 g, 94% 2 steps).

MK-108 (2.25 g, 5.56 mmol) was suspended in 90 mL of dry Et₂O. To thestirred suspension was added n-BuLi in hexane (1.6M, 4.52 mL, 7.23 mmol,1.3 eq.) and the mixture was stirred at −78° C. After 1.5 hr thesuspension turned homogeneous. The mixture was stirred for additional 30min at −78° C. Then, excessive dry CO₂ gas (ca 30 eq.) was added bybubbling through an inlet over 15 min. The resulting mixture was stirredat −78° C. for 40 min after which it was allowed to warm to rt. After 1hr the reaction mixture was quenched with saturated aqueous solution ofNa₂CO₃ and washed with Et₂O. The basic aqueous layer was acidified withKHSO₄ and extracted with AcOEt. The organic extract was washed withbrine, dried over Na₂SO₄, filtered, and concentrated to yield crude paleyellow crystals of MK-109 (1.93 g). The crude product was used for nextstep without purification.

Using similar procedure for the synthesis of MK-093 from MK-092, crudeMK-109 (1.93 g) was converted to a crude brown oil of MK-110 (1.73 g).The crude product was used for next step without purification.

Crude MK-110 (1.73 g) was dissolved in 75 mL of THF. Then, TBAF in THF(1M, 9.0 mL, 9.0 mmol, 2 eq.) was added at rt. The mixture was stirredfor 2 days before AcOEt and aqueous solution of KHSO₄C were added. Theorganic extract was washed with brine, dried over Na₂SO₄, filtered andconcentrated. The crude product was purified by chromatography on silicagel (hexane/AcOEt: 2/1) to afford colorless crystals of MK-111 (476 mg,36% 4 steps).

Using similar procedure for the synthesis of NY-90 from NY-89, MK-111(412 mg, 1.73 mmol) was converted to crude pale yellow oil of MK-112(435 mg). The crude product was used for next step without purification.

Using similar procedure for the synthesis of NY-91 from NY-90, crudeMK-112 (435 mg) was converted to colorless oil of MK-113 (494 mg, 79% 3steps).

To a stirred solution of MK-113 (423 mg, 1.17 mmol) in 9 mL of DMSO wasadded a solution of KOH (196 mg, 3.50 mmol) in 4.5 mL of water and themixture was heated to 80° C. After stirred for 2 hrs, the mixture wascooled to rt and diluted with Et₂O. The mixture was acidified withaqueous solution of KHSO₄ and extracted with AcOEt. The organic extractwas washed with brine, dried over Na₂SO₄, filtered, and concentrated togive crude pale yellow crystals of MK-114 (428 mg). It was used for nextstep without purification.

Using similar procedure for the synthesis of MK-047 from MK-046, crudeMK-114 (428 mg) was converted to pale pink oil of MK-115 (476 mg, 91% 2steps) as compound purified.

Using similar procedure for the synthesis of compound 18, theintermediate of NF2561, from compound 16, YE-06 (627 mg, 0.851 mmol)coupled with MK-115 (474 mg, 1.06 mmol) was converted to pale yellow oilof MK-116 (804 mg, 99% 3 steps).

Using similar procedure for the synthesis of YE-17 from YE-16, MK-116(800 mg, 0.844 mmol) was converted to crude oil of MK-117 (735 mg,including silyl impurity). The crude product was used for next stepwithout purification.

Using similar procedure for the synthesis of YE-18 from YE-17, the crudeMK-117 (735 mg, assumed to contain 0.844 mmol) was converted to acolorless solid of MK-118 (192 mg, 48% 3 steps).

Using similar procedure for the synthesis of 509-HD-125 from509-HD-119B, MK-118 (141 mg, 0.296 mmol) was converted to crude paleyellow solid of MK-119 (104 mg, <74%). The crude MK-119 was used fornext step without purification.

Using similar procedure for the synthesis of NF2306 from MK-090, crudeMK-119 (104 mg) was converted to crude pale yellow oil. The crudeproduct was purified by chromatography on silica gel (hexane/AcOEt: 1/2)to afford pale yellow crystals of NF2547 (59 mg, 51% 2 steps).

Synthetic Procedure for NF-2553

Using same procedure for 509-HD-209, NY-37 (4.8 g, 19.59 mmol) wasconverted to NY-117 (4.93 g).

NY-117 (2.89 g, 10 mmol) was dissolved in Et₂O (40 mL) and cooled to−78° C., under nitrogen. Then, sec-BuLi (1.3M/cyclohexane, 9.3 mL, 12.09mmol) was slowly added and the reaction was stirred at −78° C. for 30min. DMF (1.55 mL, 20 mmol) was added to the solution, then the solutionwas stirred at −78° C. for 10 min. The mixture was quenched with sat.NH₄Cl and extracted with EtOAc. The organic layer was washed with water,brine and dried over Na₂SO₄, filtered and concentrated. The crudeproduct was purified on silica gel column with hexane/EtOAc, 15:1, 10:1,8:1, 6:1, 5:1, 4:1 to give 811 mg of NY-118.

A mixture of NY-118 (4.09 g, 17.17 mmol), mCPBA (10 g, 40.56 mmol) andCHCl₃ (15 mL) was refluxed for 2.5 hrs. The reaction mixture was quencedwith sat. Na₂S₂O₃ and extracted with EtOAc. The organic layer was washedwith sat. Na₂S₂O₃, sat. NaHCO₃ (×2), brine, dried over Na₂SO₄, filteredand concentrated. The crude product was purified on silica gel columnwith hexane/EtOAc, 10:1, 8:1 to give 2.58 g of NY-119.

Using same procedure for 10, NY-119 (2.58 g, 10.15 mmol) was convertedto NY-120 (2.45 g).

A mixture of NY-120 (470 mg, 1.41 mmol), 2-bromo-1-chloroethane (0.35ml, 4.22 mmol), K₂CO₃ (292 mg, 2.11 mmol) was refluxed for 6 hrs. Then,additional 2-bromo-1-chloroethane (0.35 ml, 4.22 mmol) was added and themixture was refluxed for 15 hrs. K₂CO₃ (450 mg, 3.26 mmol) and2-bromo-1-chloroethane (1.05 ml, 12.67 mmol) were added and the mixturewas refluxed for 4 hrs. The insoluble material was filtered and thefiltrate was concentrated. The residue was diluted with EtOAc and washedwith water, brine, dried over Na₂SO₄, filtered and concentrated to give519 mg of NY-121.

A mixture of NY-121 (519 mg, 1.31 mmol), NaN₃ (212 mg, 3.26 mmol) andDMF (10 mL) was stirred at 80° C. for 3 hrs. The mixture was dilutedwith EtOAc and washed with water (×3), brine, dried over Na₂SO₄,filtered and concentrated. The crude product was purified on silica gelcolumn with hexane/EtOAc, 8:1 to give additional 452 mg of NY-122.

Using same procedure for NY-110, NY-122 (450 mg, 1.12 mmol) wasconverted to NY-123 (442 mg).

Using same procedure for 509-HD-213, NY-123 (322 mg, 0.827 mmol) wasconverted to NY-124 (365 mg).

Using same procedure for 16, YE-06 (441 mg, 0.598 mmol) was converted tocrude product of NY-125 (857 mg).

Using same procedure for 18, NY-125 (857 mg) was converted to NY-126(231 mg).

Using same procedure for 509-HD-116, NY-126 (230 mg, 0.233 mmol) wasconverted to NY-127 (197 mg). NY-127 was used without purification forthe next step.

Using same procedure for TM-12, NY-127 (197 mg, 0.233 mmol) wasconverted to NY-128 (53 mg).

To a solution of NY-128 (7.2 mg, 0.0139 mmol) in THF (1 mL), n-Bu₃P (3.5μL) was added and stirred at room temperature for 80 min. Additionaln-Bu₃P (3.5 μL) was added and the mixture was stirred for 80 min. Then,water (50 μL) was added to the reaction mixture, and the mixture wasstirred at room temperature for 3 hrs. The reaction mixture was driedover Na₂SO₄, filtered and concentrated. The crude product was purifiedon silica gel column with CH₂Cl₂/MeOH, 98:2, 95:5, 9:1 to give 5.2 mg ofNY-129.

To a solution of NY-129 (5.2 mg, 0.0106 mmol), Et₃N (5 μL, 0.0359 mmol),Ac20 (1.5 μL, 0.0159 mmol) was added at 0° C. and stirred for 40 min.The reaction mixture was quenched with sat. NH₄Cl and extracted withEtOAc. The organic layer was washed with water, sat. NaHCO₃, brine,dried over Na₂SO₄ filtered and concentrated to give 5 mg of NY-130.

Using same procedure for 509-HD-125, NY-130 (5 mg, 0.00937 mmol) wasconverted to NY-131 (6.5 mg). NY-131 was used without purification forthe next step.

Using same procedure for B2538, NY-131 (6.4 mg, 0.00937 mmol) wasconverted to NF-2553 (1.3 mg).

Synthetic Procedure for NF-2556

Using same procedure for NY-121, NY-120 (670 mg, 2 mmol) was convertedto NY-132 (875 mg).

Using same procedure for NY-122, NY-132 (873 mg, 2 mmol) was convertedto NY-133 (716 mg).

Using same procedure for NY-123, NY-133 (708 mg, 1.7 mmol) was convertedto NY-134 (694 mg).

Using same procedure for NY-124, NY-134 (694 mg, 1.7 mmol) was convertedto NY-135 (799 mg).

Using same procedure for 16, YE-06 (318 mg, 0.432 mmol) was converted tocrude product of NY-136 (683 mg).

Using same procedure for 18, NY-136 (682 mg) was converted to NY-126(229 mg).

Using same procedure for 509-HD-116, NY-137 (225 mg, 0.224 mmol) wasconverted to NY-138 (207 mg). NY-138 was used without purification forthe next step.

Using same procedure for TM-12, NY-138 (207 mg, 0.224 mmol) wasconverted to NY-139 (72 mg).

Using same procedure for NY-129, NY-139 (33 mg, 0.0621 mmol) wasconverted to NY-140 (28.5 mg).

Using same procedure for NY-130, NY-140 (28 mg, 0.0554 mmol) wasconverted to NY-141 (29.8 mg).

Using same procedure for 509-HD-125, NY-130 (5 mg, 0.00937 mmol) wasconverted to NY-131 (18 mg).

Using same procedure for B2538, NY-143 (18 mg, 0.033 mmol) was convertedto NF-2556 (12.6 mg).

Preparation of C13-C analogs, NF1774, NF2546, NF2550, NF2551, NF2552,NF2554, NF2555, and NF2560

Synthetic Procedure for NF-1774

Using same procedure for 9, NY-37 (8.1 g, 33.05 mmol) was converted toNY-38 (9.28 g).

NY-38 (31.76 g, 88.39 mmol) was dissolved in Et₂O (300 mL) and cooled to−78° C., under nitrogen. Then, t-BuLi (1.7M/pentane, 100 mL, 170 mmol)was slowly added and the reaction was stirred at −78° C. for 20 min. Dryice was added to the solution, then the solution was allowed to warm tort and was stirred for 1.5 hrs. The mixture was quenched with sat.NH₄Cl, acidified with 10% citric acid, extracted with EtOAc (×2). Theorganic layers were washed with water, brine and dried over Na₂SO₄,filtered and concentrated to give 28.39 g of NY-39.

A mixture of NY-39 (25.48 g, 78.54 mmol), dimethyl sulfate (7.8 mL,82.43 mmol), NaHCO₃ (9.9 g, 117.8 mmol) and acetone (200 mL) wasrefluxed for 14 hrs. The insoluble material was filtered and thefiltrate was concentrated. The residue was diluted with EtOAc and washedwith sat. NH₄Cl, water, brine, dried over Na₂SO₄, filtered andconcentrated. The crude product was purified on silica gel column withhexane/EtOAc, 50:1, 40:1, 30:1, 15:1 to give 7.95 g of NY-40 and 6.97 gof the desilylated product NY-41.

Using same procedure for NY-07, NY-41 (11.75 g, 52.41 mmol) wasconverted to NY-42 (13.43 g).

Using same procedure for 10, NY-42 (13.43 g, 50.44 mmol) was convertedto NY-43 (8.74 g) and the deacetylated product NY-44 (7.86 g).

A mixture of NY-43 (8.74 g, 23.34 mmol), NY-44 (7.86 g, 23.65 mmol), 2NNaOH (230 mL) and MeOH (300 mL) was refluxed for 24 hrs. The mixture wasconcentrated and the residue was acidified with 2N HCl, extracted withEtOAc. The organic layer was washed with water, brine, dried overNa₂SO₄, filtered and concentrated to give 12.62 g of NY-45.

A mixture of NY-45 (12.62 g, 41.47 mmol), conc. sulfuric acid (0.8 mL)and MeOH (200 mL) was refluxed for 15 hrs. The mixture was concentratedand the residue was diluted with EtOAc and washed with water, brine,dried over MgSO₄, filtered and concentrated. The crude product waspurified on silica gel column with hexane/EtOAc, 6:1, 5:1, 4:1, 1:1 togive 2 g of NY-46 and 11.1 g of mixture of NY-47 (small amount) andNY-48.

Using same procedure for 509-HD-213, a mixture of NY-47 and NY-48 (500mg, 1.57 mmol) was converted to NY-49 (297 mg).

Using same procedure for 509-HD-209, NY-49 (297 mg, 0.71 mmol) wasconverted to NY-50 (250 mg).

To a mixture of 60% NaH in mineral oil (330 mg, 8.25 mmol) and DMF (20mL), a solution of NY-24 (3.46 g, 5.94 mmol) in DMF (20 mL) wasgradually added at 0° C. and stirred for 30 min. Then, MPMCl (1.2 ml,8.85 mmol) was added and the reaction mixture was allowed to warm toroom temperature and stirred for 12 hrs. The reaction mixture was pouredinto ice-cooled sat. NH₄Cl and extracted with EtOAc. The organic layerwas washed with water (×2), brine, dried over Na₂SO₄, filtered andconcentrated. The crude product was purified on silica gel column withhexane/EtOAc, 18:1, 15:1 to give 1.9 g of NY-51.

Using same procedure for NY-26, NY-51 (1.9 g, 2.7 mmol) was converted toNY-52 (1.64 g).

Using same procedure for 554-RB-260, NY-52 (1.64 g, 2.65 mmol) wasconverted to NY-53 (1.68 g).

Using same procedure for 16, NY-53 (540 mg, 0.741 mmol) was converted toNY-54 (465 mg).

Using same procedure for 18, NY-54 (458 mg, 0.431 mmol) was converted toNY-55 (274 mg).

Using same procedure for 509-HD-116, NY-55 (273 mg, 0.286 mmol) wasconverted to NY-56 (166 mg).

Using same procedure for 509-HD-118, NY-56 (164 mg, 0.267 mmol) wasconverted to NY-57 (86 mg).

Using same procedure for 509-HD-188, NY-57 (30 mg, 0.05 mmol) wasconverted to NY-58 (20 mg).

To a solution of NY-58 (19 mg, 0.04 mmol) in CH₂Cl₂ (5 mL), MS4A (50 mg)and PDC (33 mg, 0.088 mmol) were added and the mixture was stirred atroom temperature for 12 hrs. The insoluble material was filtered, washedwith EtOAc and the filtrate was concentrated. The residue was dissolvedin CH₂Cl₂ (5 mL), then Dess-Martin periodinate (50 mg, 0.118 mmol) wasadded and the mixture was stirred for 4 days. Sat. NaHCO₃ and 10%Na₂S₂O₃ were added and the mixture was extracted with EtOAc. The organiclayer was washed with water, brine and dried over Na₂SO₄, filtered andconcentrated. The crude product was purified on silica gel column withhexane/EtOAc, 5:1, 4:1, 3:1, 2:1 to give 6.8 mg of NY-59.

To a stirred solution of NY-59 (6.8 mg, 0.014 mmol) in THF (0.4 mL)-H₂O(0.2 mL) was added trifluoroacetic acid (0.4 mL) at 0° C. The mixturewas then allowed to warm to rt. After 2 hrs, the mixture was poured intoa saturated solution of NaHCO₃ and extracted with EtOAc. The organiclayer was washed with water, brine and dried over anhydrous Na₂SO₄,filtered and concentrated. The residue was dissolved in MeCN (1.2 ml)and then 50% HF (0.2 mL) was added at 0° C. The mixture was then allowedto warm to rt. The crude product was purified on silica gel column withhexane/EtOAc, 1:1, 1:2 to give 2.8 mg of NF-1774.

Synthetic Procedure for NF-2546

Using same procedure for 509-HD-213, Methyl 2,5-dihydroxybenzoate (8.41g, 50 mmol) was converted to NY-99 (5.06 g).

Using same procedure for 509-HD-209, NY-99 (5.4 g, 20.91 mmol) wasconverted to NY-100 (5.83 g).

A mixture of NY-100 (5.82 g, 19.25 mmol), 10% Pd on carbon (610 mg) andMeOH (100 ml) was hydrogenated at 4 atom for 3 hrs. The catalyst wasfiltered and the filtrate was concentrated to give 4.16 g of NY-101. Itwas used without purification for the next step.

Using same procedure for 611-MS-88, NY-101 (4.15 g, 19.25 mmol) wasconverted to NY-102 (5.65 g).

To a mixture of NY-102 (5.64 g, 16.38 mmol), PhB(OH)₂ (4 g, 32.81 mmol),K₂CO₃ (3.4 g, 24.6 mmol) and toluene (120 mL) under N2 bubbling,Pd(Ph3P)₄ (570 mg, 0.493 mmol) was added and the mixture was graduallywarmed to 90° C. and stirred for 2 hrs. The reaction mixture wasfiltered and the filtrate was extracted with EtOAc. The organic layerwas washed with sat. NaHCO₃, brine, dried over Na₂SO₄, filtered andconcentrated. The crude product was purified on silica gel column withhexane/EtOAc, 20:1, 15:1, 12:1 to give 4.14 g of NY-103.

Using same procedure for NY-85, NY-103 (4.14 g, 15.2 mmol) was convertedto NY-104 (2.75 g).

Using same procedure for NY-86, NY-104 (2.7 g, 8.61 mmol) was convertedto NY-105 (822 mg).

Using same procedure for NY-87, NY-105 (810 mg, 2.37 mmol) was convertedto NY-106 (689 mg).

A mixture of NY-106 (682 mg, 1.986 mmol), CSA (22 mg, 0.095 mmol) andtoluene (15 mL) was refluxed for 4 hrs. The reaction mixture wasconcentrated, then the crude product was purified on silica gel columnwith hexane/EtOAc, 10:1, 6:1 to give 471 mg of NY-107.

Using same procedure for NY-90, NY-107 (470 mg, 1.74 mmol) was convertedto NY-108 (500 mg). NY-108 was used without purification for the nextstep.

Using same procedure for NY-91, NY-108 (470 mg, 1.74 mmol) was convertedto NY-109 (336 mg).

A mixture of NY-109 (330 mg, 0.837 mmol), KOH (142 mg, 2.53 mmol), water(5 mL) and DMSO (15 mL) was heated at 70° C. for 24 hrs. The reactionmixture was diluted with EtOAc and washed with 10% KHSO₄, water, brine,dried over Na₂SO₄, filtered and concentrated to give 334 mg of NY-110.NY-110 was used without purification for the next step.

Using same procedure for 509-HD-213, NY-110 (333 mg, 0.837 mmol) wasconverted to NY-111 (359 mg).

Using same procedure for 16, YE-06 (394 mg, 0.535 mmol) was converted tocrude product of NY-112 (687 mg).

Using same procedure for 18, NY-112 (682 mg) was converted to NY-113(215 mg).

Using same procedure for 509-HD-116, NY-113 (236 mg, 0.241 mmol) wasconverted to NY-114 (198 mg). NY-114 was used without purification forthe next step.

Using same procedure for TM-12, NY-114 (198 mg, 0.24 μmol) was convertedto NY-115 (31 mg).

Using same procedure for 509-HD-125, NY-115 (43 mg, 0.0845 mmol) wasconverted to NY-116 (47 mg).

Using same procedure for B2538, NY-116 (45 mg, 0.0845 mmol) wasconverted to NF-2546 (29.4 mg).

Synthetic Procedure for NF2548

TM-51 (1.0 g, 2.0 mmol) was dissolved in triethylamine (25 mL). MK-123(0.55 g, 3.1 mmol), PdCl₂(PPh₃)₂ (220 mg, 0.31 mmol), CuI (120 mg, 0.63mmol) were added and the mixture was stirred at 70° C., while additionof MK-123 (0.25 g, 1.4 mmol/each time) were repeated at intervals of 1hr for five times. The mixture was cooled to rt and concentrated. Theconcentrate was triturated with diethyl ether and hexane, and theinsoluble solid was filtered off. The filtrate was concentrated and theresidue was purified by chromatography on silica gel using 5-10%EtOAc/hexane to give 0.67 g (1.2 mmol, 56%) of YE-32.

Using similar procedure for the intermediate 18 of NF2561, the iodide(430 mg, 0.58 mmol) was converted to YE-33 (430 mg, 0.40 mmol, 68% 3steps).

Using similar procedure for TM-12, YE-33 (430 mg, 0.40 mmol) wasconverted to YE-34 (113 mg, 0.19 mmol, 47%, 3 steps).

Using similar procedure for TM-13, YE-34 (40 mg, 0.066 mmol) wasconverted to enone. The following deprotection of the enone by DDQresulted in deprotection of both MPM and MOM group and provided 16 mg ofYE-35 (0.036 mmol, 55% 2 steps).

Using similar procedure for YE-19, YE-35 (12 mg, 0.027 mmol) wasconverted to NF-2548 (10.5 mg, 0.026 mmol, 96%).

Synthetic Procedure for NF2549

YE-35 (4 mg, 0.0091 mmol) and triethylamine (0.01 mL, 0.071 mmol) weredissolved in THF (0.6 mL). Methyl isocyanate (total 0.070 mL, 1.2 mmol)was added in four portions in 2 h intervals at rt. After 4 days, excessisocyanate was quenched with ethyleneglycol (0.15 ml). The mixture waspoured into water and extracted with EtOAc. The extract was washed with5% KHSO₄ aq., a saturated NaHCO₃ sol., brine, dried over anhydrousNa₂SO₄, filtered and concentrated. The crude product was purified bychromatography on silica gel using 15-30% EtOAc/hexane to give 4 mg(0.0080 mmol, 88%) of the N-methylcarbamate.

Using similar procedure for ER803064, the N-methylcarbamate (4 mg,0.0080 mmol) was converted to NF2549 (3 mg, 0.0028 mmol, 35%).

Synthetic Procedure for NF2554, NF2555

Using similar procedure for YE-32, the TM-49 (2.2 g, 8.0 mmol) wasconverted to YE-36 (2.1 g, 5.8 mmol, 72%).

To a stirred solution of YE-36 (2.1 g, 5.7 mmol) in EtOH (60 mL) and THF(15 mL) was added Raney Ni W-2 (water suspension, 4.5 g) under nitrogenatmosphere. The suspension was stirred under hydrogen atmosphereovernight. Then the catalyst was removed by filtration, and the filtratewas concentrated. The concentrate was purified by chromatography onsilica gel using 15-33% EtOAc/hexane to give 1.6 g (4.3 mmol, 75%) ofYE-37.

Using similar procedure for TM-51, YE-37 (1.18 g, 3.2 mmol) wasconverted to YE-38 (0.20 g, 0.40 mmol, 13%, 3 steps).

Using similar procedure for TM-39, YE-38 (340 mg, 0.68 mmol) wasconverted to YE-39 (350 mg, 0.64 mmol, 88% 2 steps).

Using similar procedure for the intermediate 18 of NF2561, the iodide(230 mg, 0.30 mmol) was converted to YE-40 (115 mg, 0.11 mmol, 34% 3steps).

Using similar procedure for TM-12, YE-40 (115 mg, 0.11 mmol) wasconverted to YE-41 (36 mg, 0.059 mmol, 55% 3 steps).

Using similar procedure for TM-13 followed by usual deprotection of MPMgroup by DDQ oxidation, YE-41 (36 mg, 0.059 mmol) was converted to YE-42(13 mg, 0.027 mmol, 45% 2 steps).

Using similar procedure for ER803064, YE-42 (5 mg, 0.010 mmol) wasconverted to NF2554 (4.1 mg, 0.010 mmol, 99%).

Using similar procedure for NF-2549, YE-42 (7 mg, 0.014 mmol) wasconverted to NF-2555 (1.9 mg, 0.0041 mmol, 29% 2 steps).

Synthetic Procedure for NF2550

Using similar procedure for 509-HD-207, starting material (4.91 g, 25mmol) was converted to TM-34 (5.17 g, 18 mmol, 72%).

To a stirred suspension of NaH (60% oil dispersion, 593 mg, 14.8 mmol)in dry DMF (5 mL) was added a solution of TM-34 (3.395 g, 11.9 mmol) indry DMF (35 mL) at 0° C. under nitrogen atmosphere. After 30 min, methyliodide (3.7 mL, 59 mmol) was added. the mixture was allowed to warm toit and stirred for 30 min after which a saturated solution of NH₄Cl wasadded. The mixture was extracted with EtOAc and the organic extract waswashed with water, brine, dried with anhydrous Na₂SO₄, filtered andconcentrated.

The crude product (4.33 g) was dissolved in EtOH (60 mL). Then, 10% Pdon carbon (630 mg) was added. Reaction was stirred under hydrogen. After3 hrs, reaction was stopped, catalyst was filtered through celite andmixture was concentrated under reduced pressure to give the phenol (2.94g) as crude product.

To a solution of the phenol (2.94 g) in thy DMF (60 mL) were addedanhydrous K₂CO₃ (3.28 g, 24 mmol) and 2-bromo-1,1-diethoxyethane (2.86mL, 18 mmol) and the mixture was stirred for 5 hrs at 140° C. Aftercooling to rt, the mixture was poured into water and extracted withEtOAc. The organic extract was washed with a saturated solution ofNH₄Cl, water, brine, dried with anhydrous Na₂SO₄, filtered andconcentrated. The crude product was purified by chromatography on silicagel using 15% EtOAc/hexane to give 3.369 g (10 mmol, 87% 3 steps) ofTM-35.

To a solution of TM-35 (3.02 g, 9.3 mmol) in toluene (120 mL) was addedPPA (5.9 mL) and the mixture was stirred for 2 hrs at 100° C. undernitrogen atmosphere. After cooling to rt, the mixture was poured intoice/water and extracted with EtOAc. The organic extract was washed withwater, a saturated solution of NaHCO₃, brine, dried with anhydrousNa₂SO₄, filtered and concentrated. The crude product was purified bychromatography on silica gel using 12.5% EtOAc/hexane to give 395 mg(1.7 mmol, 18%) of TM-36.

To a stirred solution of TM-36 (541 mg, 2.3 mmol) in dry CH₂Cl₂ (20 mL)was added BBr₃ (0.44 mL, 4.6 mmol) at −78° C. under nitrogen atmosphere.After 1 hr, the mixture was poured into water and extracted with EtOAc.The organic extract was washed with a saturated solution of NaHCO₃,water, brine, dried with anhydrous Na₂SO₄, filtered and concentrated.The crude product was purified by chromatography on silica gel using 5%EtOAc/hexane to give 357 mg (1.6 mmol, 70%) of the phenol.

To a stirred solution of the phenol (170 mg, 0.77 mmol) in dry THF (8mL) were added DBU (0.23 mL, 1.5 mmol) and chloromethyl methyl ether(0.12 mL, 1.5 mmol) at rt under nitrogen atmosphere. After 3 hr, themixture was poured into a saturated solution of NH₄Cl and extracted withEtOAc. The organic extract was washed with 10% KHSO₄ aq., water, asaturated solution of NaHCO₃, brine, dried with anhydrous Na₂SO₄,filtered and concentrated. The crude product was purified bychromatography on silica gel using 15% EtOAc/hexane to give 193 mg (0.73mmol, 95%) of TM-37.

Using similar procedure for the intermediate 10 of NF2561, TM-37 (192mg, 0.73 mmol) was converted to TM-38 (192 mg, 0.52 mmol, 71% 2 steps).

Using similar procedure for the intermediate 14 of NF2561, TM-38 (192mg, 0.52 mmol) was converted to TM-39 (190 mg, 0.43 mmol, 83% 2 steps).

Using similar procedure for the intermediate 18 of NF2561, the iodide(223 mg, 0.30 mmol) was converted to TM-40 (232 mg, 0.25 mmol, 81% 3steps).

Using similar procedure for TM-12, TM-40 (232 mg, 0.25 mmol) wasconverted to TM-41 (81 mg, 0.17 mmol, 70% 3 steps).

Using similar procedure for ER803064, TM-41 (41 mg, 0.087 mmol) wasconverted to NF2550 (20 mg, 0.052 mmol, 60% 2 steps).

Synthetic Procedure for NF2560

To a stirred solution of 2,2,6,6-tetramethylpiperidine (0.05 mL, 0.3mmol) in dry THF (1.5 mL) was added 1.6M n-BuLi in hexane (0.15 mL, 0.24mmol) at −20° C. under nitrogen atmosphere. After 30 min, the mixturewas cooled to −78° C. and a solution of TM-41 (38 mg, 0.08 mmol) in dryTHF (4 mL) was added. After 1 hr, dry DMF (0.06 mL) was added and themixture was allowed to warm to rt. It was quenched with a saturatedsolution of NH₄Cl and extracted with EtOAc. The organic extract waswashed with a saturated solution of NH₄Cl, water, brine, dried withanhydrous Na₂SO₄, filtered and concentrated. The crude product waspurified by chromatography on silica gel using 40% EtOAc/hexane to give18 mg (0.036 mmol, 45%) of TM-42.

To a stirred solution of TM-42 (54 mg, 0.11 mmol) in CH₂Cl₂ (5 mL) wereadded triethylamine (0.3 mL, 2.2 mmol), small amount ofN,N-dimethylaminopyridine, acetic anhydride (0.1 mL, 1.1 mmol) at 0° C.The mixture was allowed to warm to rt and stirred overnight. It wasquenched with a saturated solution of NH₄Cl and extracted with EtOAc.The organic extract was washed with a saturated solution of NH₄Cl,water, brine, dried with anhydrous Na₂SO₄, filtered and concentrated.

The crude acetate (61 mg) was dissolved in MeOH (5 mL) and NaBH₄ (10 mg,0.26 mmol) was added at 0° C. After 10 min, the mixture was poured intoa saturated solution of NH₄Cl and extracted with EtOAc. The organicextract was washed with a saturated solution of NH₄Cl, water, brine,dried with anhydrous Na₂SO₄, filtered and concentrated. The crudeproduct was purified by chromatography on silica gel using 50%EtOAc/hexane to give 40 mg (0.073 mmol, 68% 2 steps) of TM-43.

To a stirred solution of TM-43 (40 mg, 0.073 mmol) in dry THF (8 mL)were added triphenylphosphine (58 mg, 0.22 mmol), diphenylphosphorylazide (0.047 mL, 0.22 mmol), 40% DEAD in toluene (0.1 mL, 0.22 mmol) atrt. After 20 hrs, the mixture was concentrated under reduced pressureand the residue was purified by chromatography on silica gel using 25%EtOAc/hexane to give 26 mg (0.046 mmol, 62%) of TM-44.

To a stirred solution of TM-44 (26 mg, 0.046 mmol) in dry THF (2.5 mL)was added tributylphosphine (0.035 mL, 0.14 mmol) at rt under nitrogenatmosphere. After 30 min, H₂O (0.5 mL) was added. After additional 3hrs, the mixture was diluted with EtOAc and dried with anhydrous Na₂SO₄,filtered and concentrated.

The crude amine (56 mg) was dissolved in CH₂Cl₂ (3 mL). Thentriethylamine (0.036 mL, 0.26 mmol) and methanesulfonyl chloride (0.010mL, 0.13 mmol) were added at 0° C. After 30 min, the mixture was pouredinto a saturated solution of NaHCO₃ and extracted with EtOAc. Theorganic extract was washed with a saturated solution of NaHCO₃, water,brine, dried with anhydrous Na₂SO₄, filtered and concentrated.

The crude methanesulfonamide (64 mg) was dissolved in EtOH (3 mL) and 1NNaOH aq. (0.5 mL, 0.5 mmol) was added at rt. After 24 hrs, the mixturewas quenched with 1N HCl (0.5 mL) and extracted with EtOAc. The organicextract was washed with water, brine, dried with anhydrous Na₂SO₄,filtered and concentrated. The crude product was purified bychromatography on silica gel using 60% EtOAc/hexane to give 15 mg (0.026mmol, 57% 3 steps) of TM-45.

Using similar procedure for ER803064, TM-45 (15 mg, 0.026 mmol) wasconverted to NF2560 (4.4 mg, 0.0089 mmol, 35% 2 steps).

Synthetic Procedure for NF2545

To a stirred suspension of 3-methoxybenzyl alcohol (10.78 g, 78 mmol) inhexane (100 mL) was added 1.6M n-BuLi in hexane (100 mL, 160 mmol) at−30° C. under nitrogen atmosphere. After 1.5 hrs, dry CO₂ was bubbledfor 30 min and then the mixture was poured into water and extracted withdiethyl ether. The aqueous layer was acidified by addition of 5N HCl at0° C. and stirred overnight at rt. The precipitate was collected byfiltration and washed with water, dried at 70° C. for 24 hrs to give4.79 g (29 mmol, 37%) of TM-46.

TM-46 (4.79 g, 29 mmol) was dissolved in CH₂Cl₂ (150 mL). Then bromine(1.88 mL, 37 mmol) was added at 0° C. and the mixture was allowed towarm to rt. After 20 hrs, the mixture was poured into a saturatedsolution of Na₂S₂O₃ and extracted with EtOAc. The organic extract waswashed with a saturated solution of NaHCO₃, water, brine, dried withanhydrous Na₂SO₄, filtered and concentrated to give 6.59 g (27 mmol,93%) of TM-47.

Using similar procedure for TM-37, TM-47 (5.37 g, 22 mmol) was convertedto TM-49 (5.98 g, 22 mmol, 99% 2 steps).

To a stirred solution of TM-49 (3.85 g, 14 mmol) in DMSO (40 mL) wasadded a solution of KOH (851 mg, 15 mmol) in H₂O (15 mL). After 30 min,H₂O was removed by evaporation and methyl iodide (2.63 mL, 42 mmol) wasadded and the mixture was stirred for 30 min. The mixture was dilutedwith EtOAc and washed with water, a saturated solution of NaHCO₃, water,brine, dried with anhydrous Na₂SO₄, filtered and concentrated.

The crude product (3.79 g) was dissolved in dry DMF (100 mL). Thendiphenyl disulfide (6.16 g, 28 mmol), pyridine (4.55 mL, 56 mmol),tributylphosphine (7.03 mL, 28 mmol) were added at rt under nitrogenatmosphere and the mixture was stirred overnight. It was quenched with asaturated solution of NaHCO₃ and extracted with EtOAc. The organicextract was washed with 10% KHSO₄ aq., water, brine, dried withanhydrous Na₂SO₄, filtered and concentrated. The crude product waspurified by chromatography on silica gel using 15% EtOAc/hexane to give1.79 g (4.5 mmol, 32% 2 steps) of TM-50.

Using similar procedure for TM-39, TM-50 (1.79 g, 4.5 mmol) wasconverted to TM-51 (1.88 g, 3.9 mmol, 87% 2 steps).

TM-51 (1.86 g, 3.8 mmol) was dissolved in DMF (30 mL). Thentriethylamine (10 mL), 3-butyn-1-ol (1.16 mL, 15 mmol), PdCl₂(PPh₃)₂(404 mg, 0.58 mmol), CuI (219 mg, 1.2 mmol) were added and the mixturewas stirred overnight at 70° C. The mixture was diluted with 50%EtOAc/hexane and washed with water, brine, dried with anhydrous Na₂SO₄,filtered and concentrated. The crude product was purified bychromatography on silica gel using 30% EtOAc/hexane to give 1.76 g (3.7mmol, 97%) of TM-52.

Using similar procedure for TM-45, TM-52 (754 mg, 1.6 mmol) wasconverted to TM-53 (586 mg, 1.1 mmol, 67% 3 steps).

Using similar procedure for TM-39, the iodide (350 mg, 0.48 mmol) wasconverted to TM-54 (224 mg, 0.25 mmol, 45% 3 steps).

Using similar procedure for NF2550, TM-54 (224 mg, 0.21 mmol) wasconverted to NF2545 (2.8 mg, 0.0057 mmol, 2.7% 5 steps).

Synthetic Procedure for NF2551 and NF2552

Using similar procedure for TM-52 followed by hydrogenation, TM-49 (2.95g, 11 mmol) was converted to TM-55 (2.41 g, 9.1 mmol, 84% 2 steps).

To a stirred solution of TM-55 (2.41 g, 9.1 mmol) in cyclohexane (30mL)-CH₂Cl₂ (15 mL) were added CSA (105 mg, 0.45 mmol) and4-methoxybenzyl trichloroacetimidate (7.68 g, 27 mmol) at rt undernitrogen atmosphere. After 14 hrs, triethylamine (0.1 mL) was added.Then precipitate was removed by filtration and washed with hexane. Thefiltrate was concentrated under reduced pressure. The residue waspurified by chromatography on silica gel using 30% EtOAc/hexane to give3.26 g (8.4 mmol, 93%) of TM-56.

Using similar procedure for TM-50, TM-56 (3.26 g, 8.4 mmol) wasconverted to TM-57 (1.78 g, 3.5 mmol, 41% 3 steps).

Using similar procedure for TM-39, TM-57 (366 mg, 0.72 mmol) wasconverted to TM-58 (381 mg, 0.64 mmol, 89% 2 steps).

Using similar procedure for TM-39, the iodide (223 mg, 0.30 mmol) wasconverted to TM-59 (132 mg, 0.12 mmol, 40% 3 steps).

Using similar procedure for TM-12, TM-59 (132 mg, 0.12 mmol) wasconverted to TM-60 (32 mg, 0.051 mmol, 43% 3 steps).

Using similar procedure for TM-13 followed by usual deprotection of MPMgroup by DDQ oxidation, TM-60 (32 mg, 0.051 mmol) was converted to TM-61(15 mg, 0.030 mmol, 58% 2 steps).

Using similar procedure for ER803064, TM-61 (8 mg, 0.016 mmol) wasconverted to NF2551 (4 mg, 0.010 mmol, 60%).

To a stirred solution of TM-61 (7 mg, 0.014 mmol) in CH₂Cl₂ (1 mL) wereadded triethylamine (0.02 mL) and methyl isocyanate (0.1 mL) at rt.After 15 hrs, the mixture was diluted with EtOAc and washed with 3%NH₄OH aq., 5% KHSO₄ aq., water, brine, dried with anhydrous Na₂SO₄,filtered and concentrated. The crude product was purified bychromatography on silica gel using 55% EtOAc/hexane to give 7 mg (0.013mmol, 90%) of the N-methylcarbamate.

Using similar procedure for ER803064, the N-methylcarbamate (7 mg, 0.013mmol) was converted to NF2552 (3 mg, 0.0063 mmol, 50%).

Preparation of the Intermediate for C14-O-Linked Including C₂-C₄Analogs:

Known compound 1 (0.22 g, 1.2 mmol) and K₂CO₃ (0.25 g, 1.8 mmol) wasdissolved in 12 mL of DMF. After the addition of MPM-Cl (0.17 mL, 1.2mmol) the mixture was heated at 35° C. for 12 hours. Crude mixture wasconcentrated and filtered. The resulted material was purified by silicagel chromatography. CH₂Cl₂ was used to recover 445-ycs-252 (0.16 g, 0.53mmol) from silica gel plug in 44% yield.

60% of NaH in mineral oil (16 mg, 0.40 mmol) was added to 445-ycs-252(36 mg, 0.12 mmol) in 2 mL of DMF. After the addition of MOMCl (0.17 pt,0.22 mmol), the mixture was stirred at rt for 1 hour. DMF was evaporatedunder high vacuum, and the residue was dissolved in CH₂Cl₂, washed withH₂O. The crude material was purified by silica gel chromatography. 30%EtOAc in hexanes was used to elute 445-ycs-254 (40 mg, 0.12 mmol) fromsilica gel plug in 97% yield.

2.5 M n-BuLi in hexanes (13 mL, 33 mmol) was introduced drop wise to thestirring solution of diisopropylamine (4.6 mL, 33 mmol) in 30 mL of THFat −5° C. The solution was stirred at 0° C. for 30 minutes before it wascooled to −78° C. 445-ycs-254 (6.3 g, 18 mmol) in 25 mL of THF was addedto the cold LDA slowly so that the internal temperature was kept below−78° C. The mixture was stirred at −78° C. for 45 minutes, and then(PhSe)₂ (5.2 g, 17 mmol) in 25 mL of THF was added slowly so that theinternal temperature was kept below −60° C. Reaction mixture was stirredfor 40 minutes before it was quenched with aq. NH₄Cl at −78° C. EtOAcwas added to the mixture at rt. After separation, organic layer wasdried (Na₂SO₄) and concentrated. 5% EtOAc in toluene was used to elute445-ycs-268 (6.9 g, 14 mmol) from silica gel column in 75% yield.

2.5 N NaOH (18 mL, 46 mmol) was added to 445-ycs-268 (7.7 g, 15 mmol) in18 mL of EtOH. The mixture was refluxed for 12 hours before it wasacidified and extracted with EtOAc. Organic phase was dried (Na₂SO₄) andconcentrated to afford white crystalline 445-ycs-272 (7.2 g, 15 mmol) in96% yield.

DEAD (3.5 mL, 22 mmol) was added to a solution of 445-ycs-272 (7.2 g, 15mmol), Ph₃P (5.8 g, 22 mmol) and 2-TMS-ethanol (2.6 mL, 18 mmol) in 200mL of toluene at 0° C. The mixture was stirred at RT for 1 hour beforeit was quenched with aq. NaHCO₃. Organic phase was dried (Na₂SO₄),concentrated and purified by silica gel chromatography to afford445-ycs-273 (8.2 g, 14 mmol) in 95% yield.

A procedure similar to the preparation of 554-RB-260 was used.

LiHMDS in THF (2.8 mL, 2.8 mmol) was introduced drop wise into a cold(−78° C.) solution of 445-ycs-274 (1.4 g, 1.9 mmol) and 445-ycs-273 (1.7g, 2.8 mmol) in 10 mL of 10:1 THF-HMPA. The internal temperature waskept below −70° C. during the addition. The mixture was stirred at −78°C. for half hour before it was quenched with aq. NH₄Cl and diluted withEtOAc. Organic phase was dried (Na₂SO₄), concentrated and purified bysilica gel chromatography to afford 445-ycs-278 (1.9 g, 1.6 mmol) in 82%yield.

m-CPBA (1.0 g, 4.2 mmol) was added in three portions into a cold (0° C.)solution of 445-ycs-278 (2.5 g, 2.1 mmol) in 30 mL of CH₂Cl₂. Themixture was stirred at 0° C. for 1 hour before the addition of Et₃N (1.8mL, 13 mmol). After the reaction mixture was stirred at rt for 1 hour itwas quenched with aq. Na₂S₂O₃ and diluted with aq. NaHCO₃. Organic phasewas dried (Na₂SO₄), concentrated and purified by silica gelchromatography to afford 445-ycs-281 (1.6 g, 1.6 mmol) in 76% yield.

In a solution of TBAF buffered with 0.33 mole equivalent ofimidazole.HCl (1.3 mL, 1.3 mmol) was introduced to the solution of445-ycs-281 (0.17 g, 0.17 mmol) in 2 mL of THF. The mixture was stirredat 50° C. for 12 hours before it was diluted with Et₂O and washed withaq. NH₄Cl. Organic phase was dried (Na₂SO₄) and concentrated to furnishcrude 445-ycs-295, which dissolved in 20 mL of CH₂Cl₂ was added dropwise to the refluxing mixture of 2-chloro-1-methylpyridinium iodide(0.12 g, 0.48 mmol) and n-Bu₃N (0.11 mL, 0.48 mmol) in 12 mL of CH₂Cl₂.The mixture was refluxed for 2 hours before it was diluted with Et₂O andwashed with 0.05 N HCl, H₂O and NaHCO₃. Organic phase was dried(Na₂SO₄), concentrated and purified by silica gel chromatography toafford 445-ycs-299 (90 mg, 0.13 mmol) in 81% yield for 2 steps.

p-TsOH.H₂O (11 mg, 0.060 mmol) was added in two portions to a solutionof 445-ycs-299 (40 mg, 0.060 mmol) in 4.5 mL of 2:1 MeOH-THF at rt. Thesolution was stirred at 40° C. for 3 days before it was concentrated andpurified by silica gel chromatography to afford 560-ycs-30 (15 mg, 0.032mmol) in 53% yield.

Catalytic amount of p-TsOH.H₂O (1 crystal) was added to the solution of560-ycs-30 (13 mg, 0.028 mmol) and excess amount of 2-methoxypropane (2drops) in 1 mL of CH₂Cl₂. The mixture was stirred at RT for 2 hoursbefore the addition of NaHCO₃ and filtration. The solution wasconcentrated and purified by silica gel chromatography to afford560-ycs-36 (11 mg, 0.022 mmol) in 79% yield.

Preparation of ER804104:

560-ycs-36 was used as an advanced intermediate for following analogssynthesis as examples.

1 N NaOH (2.0 mL, 2.0 mmol) was introduced into a solution of445-ycs-299 (43 mg, 0.064 mmol) in 3 mL of 2:1 EtOH-THF at rt. Thesolution was stirred at 40° C. for 12 hours before it was diluted withEt₂O and brine. Organic phase was dried (Na₂SO₄) and purified by silicagel chromatography to afford 445-ycs-311 (33 mg, 0.058 mmol) in 91%yield.

PCC (38 mg, 0.17 mmol) was added in three portions to a mixture of445-ycs-311 (33 mg, 0.058 mmol), 4 Å molecular sieves (40 mg) and celite(40 mg) in 2 mL of CH₂Cl₂ at rt. The mixture was stirred at rt for 1hour before it was diluted with Et₂O and filtered. The filtrate wasconcentrated and passed through a short plug of silica gel (1:1EtOAc-Hexanes) to afford 560-ycs-9 (28 mg, 0.049 mmol) in 86% yield.

560-ycs-9 was deprotected as described for the synthesis of ER803064 togive ER804104.

ER-804168

The synthesis of ER-804168 was the same as the synthesis of ER-803064

Synthesis of ER-805125:

The synthesis of 507-XYL-147 from 507-XYL-111 was followed the sameprocedures as in the synthesis of ER-803064.

To 507-XYL-147 (1.43 g, 1.29 mmol) in a co-solvent of THF/water (5:1v/v, 60 mL) at room temperature was added OXONE and the mixture wasstirred at room temperature for six hours. The mixture was diluted withethyl acetate and washed three times with water. The organic layer wasdried (sodium sulfate), concentrated and purified by silica gelchromatography to obtain 1.05 g (86%) of the desired product,507-XYL-148 which was confirmed by NMR and MS (M+Na=972).

The synthesis of 507-XYL-154 from 507-XYL-148 was followed the sameprocedures as in the synthesis of ER-803064.

To 507-XYL-154 (340 mg, 0.57 mmol) in a co-solvent of THF/water (4:1v/v, 12.5 mL) at 0° C. were added N-methyl morpholino-N-oxide (82.5 mg,0.68 mmol) and then osmium tetraoxide in toluene (0.1 M, 0.6 mL, 0.06mmol) in portions. The mixture was stirred at 0° C. for 19 hours. Thereaction was quenched with saturated sodium thiosulfate solution anddiluted with ethyl acetate. The mixture was washed with saturated sodiumthiosulfate solution and water. The aqueous layers were back extractedtwice with ethyl acetate and three times with methylene chloride. Thecombined organic layers were dried (sodium sulfate), concentrated andpurified by preparative TLC eluting with 10% methanol in methylenechloride to yield 158 mg (44%) of desired product, 507-XYL-165 which wasconfirmed by NMR and MS (M+Na=649).

To 507-XYL-165 (120 mg, 0.197 mmol) were added methylene chloride (5mL), 2,2-dimethoxypropane (0.2 mL, 1.63 mmol) and pyridinium tosylate(12 mg, 0.048 mmol). The mixture was stirred at room temperature for twohours. The mixture was diluted with methylene chloride and washed twicewith saturated sodium bicarbonate solution. The organic layer wasconcentrated and purified by prep TLC to obtain 131 mg of the desiredproducts, 507-XYL-168 which were confirmed by NMR and MS (M+Na=689).

The remaining synthesis of ER-805125 from 507-XYL-168 was followed thesame procedures as in the synthesis of ER-803064.

Preparation of ER-805216 and ER-805217:

The starting material 507-XYL-165 (42 mg, 0.066 mmol) was azeotropedwith toluene and dried under high vacuum for one hour. The startingmaterial was then dissolved in dry methylene chloride (5 mL) and cooledto 0° C. To this were added collidine (20.3 uL, 0.15 mmol) and thenadded methanesulfonyl anhydride solution in methylene chloride (0.1 M,0.69 mL, 0.069 mmol) drop wise over five minutes. The mixture wasstirred at 0° C. for 1.5 hour and at 4° C. over night. The mixture waspoured into a saturated sodium bicarbonate solution and extracted threetimes with methylene chloride and once with ethyl acetate. The combinedorganic layers were dried (sodium sulfate), concentrated and purified bypreparative TLC eluting with 5% methanol in methylene chloride toprovide 36 mg (77%) of the desired product, 507-XYL-178 which wasconfirmed by NMR and MS (M+Na=727).

To 507-XYL-178 (36 mg, 0.05 mmol) were added 2.0M ammonia in methanol(16 mL) and concentrated aqueous ammonium hydroxide solution (3.2 mL)and the mixture was stirred at room temperature for 12 hours. Themixture was concentrated in vacuum, azeotroped with ethyl acetate,methanol and toluene, and dried under reduced pressure. The crudeproduct, 507-XYL-192 was directly used for the next reaction.

To 507-XYL-192 in methylene chloride (10 mL) at 0° C. were addedtriethylamine (0.2 mL, 1.51 mmol) and acetic anhydride (0.1 mL, 1.06mmol). The mixture was stirred at room temperature over night. Thereaction mixture was cooled to 0° C. and quenched with saturated sodiumbicarbonate solution. The mixture was poured into excessive sodiumbicarbonate solution and extracted three times with methylene chlorideand three times with ethyl acetate. The combined organic layers weredried (sodium sulfate), concentrated to obtain the crude desiredproduct, 507-XYL-194 which was confirmed by MS (M+Na=732). The crudeproduct was directly used for the next reaction without furtherpurification.

The conversion of 507-XYL-194 to 507-XYL-198 was the same as in thesynthesis of ER-803064. Preparative TLC purified the crude product with5% ethanol in ethyl acetate to give 13.6 mg (57% over three steps) ofthe desired product, 507-XYL-198 which was confirmed by NMR and MS(M+Na=586).

The oxidation of 507-XYL-198 using pyrridinium chlorochromate wasfollowed the same procedure as in the synthesis of ER-803064.Preparative TLC purified the crude material with 8% ethanol in ethylacetate to afford 507-XYL-204a and 507-XYL-204b, which were confirmed byNMR.

ER-805216

The conversion of 507-XYL-204a to ER-805216 was the same as in thesynthesis of ER-803064. Purification of crude material by preparativeTLC with 25% ethanol in ethyl acetate afforded ER-805216, which wasconfirmed by NMR and MS (M+Na=498).

The conversion of 507-XYL-204b to ER-805217 was the same as in thesynthesis of ER-803064. Purification of crude material by preparativeTLC eluting with 25% ethanol in ethyl acetate afforded ER-805217, whichwas confirmed by NMR and MS (M+Na=500).

Preparation of ER804401:

Procedure for the preparation of 560-ycs-86 was similar to that of445-ycs-273.

Procedure for the preparation of 560-ycs-88 was similar to that of445-ycs-254.

Procedure for the preparation of ER804401 from 560-ycs-88 was similar tothe synthesis of ER804104.

ER804504 was prepared in the similar way as ER804401 from 560-ycs-36 andn-butanol.

Preparation of ER804555 and ER804567:

Procedure for the preparation of ER804555 from 560-ycs-36 and N-acetylethanolamine was similar to the synthesis of ER804401.

ER804567 was prepared in the similar way as ER804401 from 560-ycs-36 and3,4-dichlorobenzyl alcohol.

Preparation of ER804606:

ER804606 was prepared in the similar way as ER804401 from 560-ycs-36 and4-(2-hydroxyethyl)-morpholine except that oxidation of alcohol560-ycs-128 into enone 560-ycs-133 was carried out under Swernconditions as below instead of using PCC.

(COCl)₂ (2.6 μL, 0.030 mmol) was introduced into a solution of DMSO (4.2μL, 0.060 mmol) in 0.2 mL of CH₂Cl₂ at −78° C. The mixture was stirredat −78° C. for 15 minutes before the addition of 560-ycs-128 (6.0 mg,0.010 mmol) in 0.8 mL of CH₂Cl₂. After the solution was stirred for 1hour at −78° C. Et₃N (12 μL, 0.090 mmol) was added. The reaction mixturewas allowed to warm up to −20° C. for 15 minutes and quenched with Sat.NH₄Cl. Organic phase was washed with NaHCO₃ and concentrated to furnish560-ycs-133 (6.0 mg, 0.010 mmol) in quantitative yield.

Preparation of ER804630:

ER804630 was prepared in the similar way as ER804401 from 560-ycs-36 andN-acetyl, N-methyl ethanolamine.

Preparation of ER804778:

ER804778 was prepared in the similar way as ER804401 from 560-ycs-36 and3-methylthio-1-propanol except that 560-ycs-199 was oxidized into560-ycs-200 by m-CPBA.

MCPBA (12 mg, 0.048 mmol) was added in three portions to the solution of560-ycs-199 (5.2 mg, 0.0081 mmol) in 2 mL of CH₂Cl₂ at 0° C. Thesolution was stirred at 0° C. for 10 minutes before it was washed withNa₂S₂O₃ and concentrated to afford 560-ycs-200 (5.0 mg, 0.0074 mmol) in92% yield.

Aq. HCl (3.0 mL, 3.0 mmol) was introduced to a solution of 560-ycs-9 (31mg, 0.055 mmol) in 3 mL of CH₃CN at rt. The solution was stirred at rtfor 12 hours before it was diluted with EtOAc and H₂O. Organic phase waswashed with NaHCO₃, dried (Na₂SO₄), concentrated and purified by silicagel chromatography to afford ER804104 (12 mg, 0.025 mmol) in 45% yieldand ER804131 (5.9 mg, 0.016 mmol) in 30% yield.

Alternative preparation of commonly used intermediate for C14modification:

Procedure for the preparation of 560-ycs-218 was similar to that of445-ycs-273.

MOM-Cl (2.8 mL, 37 mmol) was added to a solution of 560-ycs-218 (2.1 g,7.4 mmol) and DBU (7.1 mL, 48 mmol) in 10 mL of CH₂Cl₂ at 0° C. Themixture was stirred at rt for 15 minutes before the organic phase waswashed with NaHCO₃, concentrated and purified by silica gelchromatography to furnish 560-ycs-243 (1.8 g, 5.5 mmol) from silica gelplug in 74% yield.

Procedure for the preparation of 560-ycs-248 was similar to that of445-ycs-268.

Procedure for the preparation of 560-ycs-250 was similar to that of445-ycs-272.

Procedure for the preparation of 560-ycs-251 was similar to that of445-ycs-273.

Procedure for the preparation of 560-ycs-256 was similar to that of445-ycs-278.

Procedure for the preparation of 560-ycs-258 was similar to that of445-ycs-281.

Procedure for the preparation of 560-ycs-267 was similar to that of445-ycs-295.

Procedure for the preparation of 560-ycs-269 was similar to that of445-ycs-299.

TBAF in THF (7.0 mL, 7.0 mmol) was introduced to the solution of560-ycs-269 (0.87 g, 1.3 mmol) in 7 mL of THF. The mixture was stirredat rt for 4 hours before it was diluted with Et₂O and washed with brine.Organic phase was dried (Na₂SO₄), concentrated and purified by silicagel chromatography to afford 560-ycs-270 (0.43 g, 0.78 mmol) in 58%yield over 3 steps.

Preparation of ER805190:

Transformations from 560-ycs-279 to 560-ycs-286 and from 560-yes-297 to560-ycs-300 were similar as the procedure for the preparation ofER804606.

Transformations from 560-ycs-286 to 560-ycs-297 and from 560-ycs-300 toER805190 were similar as the procedure for the preparation of ER805135.

Preparation of C14-C2 Linked Series:

To a solution of starting diphenol (10.7 g) in acetone (100 mL),BrCH₂CH₂Cl (15 mL) and 20 g of K₂CO₃ were added. The mixture was heatedat 80° C. for 1 day. It was cooled and filtered. The filtrate wasdiluted with EtOAc, washed with brine, dried and concentrated. The crudeproduct was purified on silica gel column with hexanes/CH₂Cl₂, 2:1 then1:1 to hexanes/EtOAc, 1:1 to give 12.7 g of desired product.

The MOM protection was carried out under the same conditions aspreviously described. After purification 12.7 g of product was obtained.

The mixture of chlorophenol (12.7 g), NaN₃ (6 g) in 20 mL of DMF washeated at 70° C. overnight. After cooled, it was diluted with EtOAc,washed with water, brine, dried and concentrated. The crude product waspurified on silica gel column to give 10.3 g of desired azide.

All above transformations were carried out under the conditionsidentical to previous series.

Preparation of ER804730:

ER804730 was prepared in the similar way as ER804104 from 560-ycs-171.

Preparation of ER805135:

2-Imidazole-carboxyaldehyde (96 mg, 1.0 mmol) in 0.5 mL of DMSO wasintroduced to a mixture of 560-ycs-183 (0.30 g, 0.50 mmol) and 4 Amolecular sieves (0.30 g) in 5 mL of CH₂Cl₂. The mixture was stirred for30 minutes at rt after the addition of AcOH (28 μL 0.50 mmol). K₂CO₃solid (0.14 g, 1.0 mmol), celite (0.5 g) and 10 mL of Et₂O were addedand the mixture was filtered. The filtrate was concentrated before itwas dissolved in 5 mL of EtOH and treated with NaBH₄ at RT for 5minutes. The mixture was then diluted with brine and EtOAc. Organiclayer was dried and concentrated to furnish 560-ycs-275 (0.27 g, 0.40mmol) in 80% yield. MS: 676 (M⁺+1, 100%).

Transformations from 560-ycs-275 to 560-ycs-276 and 560-ycs-277 to560-ycs-279 were similar as the procedure for the preparation ofER804104.

Boc₂O (0.27 g, 1.2 mmol) was added to a solution of 560-ycs-276 (0.29 g,0.50 mmol) and Et₃N (0.21 mL, 1.5 mmol) in 5 mL of CH₂Cl₂. The mixturewas stirred at rt for 12 hours before it was concentrated and purifiedby silica gel chromatography to provide 560-ycs-277 (0.34 g, 0.44 mmol)in 88% yield. MS: 794 (M⁺+Na, 100%).

A solution of 560-ycs-279 (0.22 g, 0.29 mmol) in 2 mL of TFA and 2 mL ofCH₂Cl₂ was sat at rt for 30 minutes before it was concentrated anddissolved in 2 mL, of CH₃CN and 2 mL of 1 N HCl. The solution wasdiluted with 10:1 CHCl₃-MeOH after it was stirred at rt for 12 hours.Organic phase was washed with NaHCO₃, concentrated and purified bysilica gel chromatography to afford ER-805135 (60 mg, 0.12 mmol) in 25%overall yield for 6 steps.

Preparation of ER804744:

Ph₃P (35 mg, 0.14 mmol) was added to a solution of 560-ycs-171 (28 mg,0.045 mmol) in 3 mL of 2:1 THF-H₂O. The solution was stirred for 12hours at it before all the volatile was evaporated and the crude residuewas dissolved in 2 mL of CH₂Cl₂. Et₃N (44 μL, 0.32 mmol) and MsCl (17μL, 0.22 mmol) was added at 0° C. The mixture was stirred at 0° C. for10 minutes before it was washed with NaHCO₃ and concentrated to provide560-ycs-184 (21 mg, 0.031 mmol) in 69% for 2 steps.

ER804744 was prepared in the similar way as ER804101 from 560-ycs-184.

Preparation of ER804759:

ER804759 was prepared in the similar way as ER804606 from 560-ycs-36 and2-(N,N-dimethylamino)-1-ethanol.

Preparation of 4 Carbon Linker C (14) Common Intermediate

491-HAD-185I

Chlorobutan-1-ol (15.5 g, 143 mmol) and DEAD (28.8 g, 165 mmol) wereadded simultaneously over 45 min to a cooled (0° C. ice/water bath)solution of diphenolic toluide (20.0 g, 110 mmol) in THF (175 mL) andtoluene (700 mL) under a nitrogen atmosphere. Then, the ice/water bathwas removed and the reaction mixture was allowed to warm to rt. Afterstirred for 2.5 h, standard Mitsunobu workup conditions followed byflash chromatography (silica gel, 5-10% EtOAc/hexanes) yielded491-HAD-185 as a colorless solid (15.66 g, 52%)

491-HAD-191

DBU (59.0 g, 388 mmol) was added drop wise to a cooled (salt/ice bath)solution of 491-HAD-185 (14.4 g, 52.9 mmol) DMF (200 mL under nitrogenatmosphere followed by chloromethyl methyl ether (26.7 g, 332 mmol).After stirred for 0.5 h, water (200 mL) was added and the aqueous phasewas extracted with CH₂Cl₂. The combined CH₂Cl₂ extracts were dried withanhydrous Na₂SO₄ and solvent was removed by reduced pressure. Flashchromatography (silica gel, 15% EtOAc/hexanes) gave 491-HAD-191 ascolorless oil (13.9 g, 81%).

491-HAD-192

Sodium azide (8.64 g, 133 mmol) was added to a solution of 491-HAD-191in DMF (150 mL) and the suspension was heated to 80° C. After stirring 2h water (200 mL) and CH₂Cl₂ (150 mL) was added. The aqueous phase wasextracted several times with CH₂Cl₂, dried with anhydrous Na₂SO₄ andsolvent removed under reduced pressure. Flash chromatography (silicagel, 20% EtOAc/hexanes) afforded 491-HAD-192 as pale yellow oil (12.9 g,90%).

491-HAD-194

Lithium diisopropylamine was prepared under nitrogen atmosphere in usualmanner from diisopropylamine (12.8 mL, 91.5 mmol) and n-butyllithium in5% HMPA/THF (150 mL) in flask equipped with overhead mechanicalstirring. LDA solution was cooled to −78° C. (dry ice/acetone bath).Next, a solution of 491-HAD-192 in 5% HMPA/THF (35 mL) was added. Afterstirring 20 min, a solution of diphenyl diselenide (12.4 g, 39.7 mmol)in 5% HMPA/THF (35 mL) was added. Small amount of intermediate wasobserved and additional solvent (20 mL) was added in an attempt tocreate a homogeneous solution. After stirred for 2.5 h, aqueous NH₄Clwas added and the aqueous phase was extracted several times with EtOAc.Combined organic extracts were washed with brine, dried with anhydrousNa₂SO₄ and solvent was evaporated under reduced pressure. Slightlyimpure 491-HAD-194 was obtained as a yellow oil (3.95 g, 21%) followingflash chromatography (silica gel, 20-100% hexanes/CH₂Cl₂, 2%EtOAc/CH₂Cl₂).

491-HAD-196

A solution of 491-HAD-194 (3.94 g, 8.24 mmol) in ethanol (200 proof,24.5 mL) and 2.5 N NaOH (16.5 mL, 41.3 mmol) was heated to 60° C. Afterstirred for 2 days, the reaction solution was then diluted with waterand hexanes and the aqueous layer was acidified with NaHSO₃ andextracted with EtOAc. The combined organic extracts were dried withanhydrous Na₂SO₄ and evaporation of the solvent under reduced pressureto afford 491-HAD-196 (3.71 g) as a pale yellow solid. 491-HAD-196 wasnot purified and was put directly into next step.

491-HAD-200

A flask equipped with nitrogen inlet was charged with 491-HAD-196 (3.70g, 7.97 mmol), CH₂Cl₂ (56 mL), DCC (6.58 g, 31.9 mmol), DMAP (97.3 mg,0.797 mmol) and triethylamine (4.44 mL, 31.9 mmol). After stirringseveral minutes 2-(trimethylsilyl)ethanol was added and the reactionmixture was heated to 35° C. for 3 days then stirred an additional 10-15h at rt. Toluene (100 mL) was added and the reaction mixture wasfiltered. The filtrate was washed with saturated aqueous NaHCO₃ solutionfollowed by brine, dried with anhydrous Na₂SO₄ and solvent was removedunder reduced pressure. Flash chromatography (silica gel, 20%EtOAc/hexanes) afforded 491-HAD-200 (1.89 g, 42%) as a colorless oil.

491-HAD-230

A solution containing iodide 554-RB-260 (1.04 g, 1.72 mmol) and491-HAD-200 (2.53 mmol) in 5% HMPA/THF (2.53 mL), under nitrogenatmosphere, was cooled to −78° C. (dry ice/acetone bath) and thereaction vessel was shielded from light. 1M LiHMDS in THF (2.53 mL, 2.53mmol) was added over 75 min by syringe pump. After stirred for anadditional 40 min at −78° C., aqueous NH₄Cl was added and the aqueousphase was extracted several times with EtOAc. Combined organic extractswere washed with brine, dried with anhydrous Na₂SO₄ and solventevaporated under reduced pressure. Flash chromatography (silica gel,5-10-15% EtOAc/hexanes) gave slightly impure (small amount of491-HAD-200) 491-HAD-230 (1.86 g) of as a colorless oil.

491-HAD-232

A solution of 491-HAD-230 (1.89 g, 1.82 mmol) in CH₂Cl₂ (26 mL) under anitrogen atmosphere, was cooled to 0° C. (ice/water bath). Next m-CPBA(57% 1.65 g, 5.46 mmol) was added in one portion. After stirred for 1.5h, triethylamine was added and ice/water bath was removed. After stirredfor an addition hour at rt, the reaction mixture was cooled withice/water bath and a solution composed of 10% v/v Na₂S₂O₃ (aqueous,saturated)/NaHCO₃ (aqueous, saturated) was added. The aqueous phase wasextracted several times with CH₂Cl₂. Combined CH₂Cl₂ extracts werewashed with saturated NaHCO₃, dried with anhydrous Na₂SO₄ and solventwas evaporated under reduced pressure. Hash chromatography of theresidue (silica gel, 15% EtOAc/hexanes) gave 491-HAD-232 as pale yellowoil (0.96 g 63% over two steps).

491-HAD-235

To a solution of 491-HAD-232 (0.96 g, 1.1 mmol) in THF (5 mL), undernitrogen atmosphere, a TBAF (1M/THF) solution (5.45 mL, 5.45 mmol),buffered with imidazole HCl (0.14 g, 1.3 mmol) was added and thereaction solution was allowed to stir at rt. After stirred for 4 days,additional TBAF (1M in THF) (2.2 mL, 2.2 mmol) was introduced to thereaction flask and heating was increased to 50° C. After stirred for 15h, heating was stopped and cooled to it, a saturated aqueous solution ofNH₄Cl was added. The aqueous phase was extracted several times withEtOAc. Combined organic phase was washed with brine, dried withanhydrous Na₂SO₄ and solvent was removed under reduced pressure. Thecrude residue was used for next step without purification.

491-HAD-237

To a solution containing 2-chloro-1-methylpyridium iodide (1.65 g, 6.47mmol), CH₂Cl₂ (80 mL) and tributylamine (1.54 mL, 6.47 mmol), heated toreflux under a nitrogen atmosphere, a solution of crude 491-HAD-235 indichloromethane (160 mL) was added over 3.5 h using syringe pump. Afterstirred for an additional hour, the heat was shut off and the reactionwas then allowed to stir at rt overnight. The reaction solution then wasconcentrated under reduced pressure and the residue was diluted withEtOAc and water. The aqueous phase was extracted several times withEtOAc and the combined organic phase was washed with 0.05 M HCl (3×85mL), saturated aqueous NaHCO₃, brine successively, dried with Na₂SO₄ andconcentrated down under reduced pressure. Flash chromatography of theresidue (silica gel, 30% ethyl acetate) afforded 491-had-237 as a paleyellow gel (0.46 g, 65% over two steps).

Preparation of Imidazole C.14 Analogue, ER805023:

491-HAD-251

A solution of 491-HAD-237 (71.4 mg, 0.110 mmol), triphenylphosphine(86.5 mg, 0.330 mmol) in THF (2 mL) and water (1 mL) was stirred at rt.After stirred for approximately 17 h, the reaction solution wasconcentrated under reduced pressure. The residue was azeotroped severaltimes with toluene and used directly in the next reaction withoutpurification.

491-HAD-252

To a flask charged with 491-HAD-254 (0.110 mmol) and molecular sieves (4A) in CH₂Cl₂, 2-imidazolecarboxaldehyde (21.1 mg, 0.220 mmol) in hotDMSO (0.30 mL) was added followed by acetic acid (6.3 μL). After stirredfor 45 min at rt, anhydrous K₂CO₃ (0.030 g) was added. The reactionmixture was subsequently diluted with diethyl ether and filtered.Following the removal of solvent, the residue was re-dissolved inmethanol and resulting solution cooled with ice/water bath and NaBH₄(0.020 g, 0.53 mmol) was added. After stirred for 20 min, brine solutionwas added and the aqueous phase was extracted several times with EtOAc.Combined EtOAc extracts were dried with anhydrous Na₂SO₄ andconcentrated. The crude residue was not purified and was used directlyin next reaction.

491-HAD-254

A solution containing 491-HAD-252 (0.110 mmol), 1 N NaOH (3.30 mL, 3.30mmol), ethanol (3.3 mL) and THF (1.6 mL) was stirred at 40-45° C. After16 h, water was added followed by CH₂Cl₂. The aqueous phase wasextracted several times with CH₂Cl₂ and the combined organic phases weredried with anhydrous Na₂SO₄ and concentrated. The crude residue was notpurified and was used directly in next reaction.

491-HAD-256

A solution of 491-HAD-254 (0.110 mmol), triethylamine (46.0 μL, 0.330mmol) and Boc anhydride (60.0 mg, 0.275 mmol) in CH₂Cl₂ was stirredunder a nitrogen atmosphere at rt for 4 days. Reaction mixture wasconcentrated under reduced pressure. Flash chromatography (solventgradient used: 20% EtOAc/hexanes, 30% EtOAc/hexanes, 50% EtOAc/hexanes,75% EtOAc/hexanes) yielded 491-HAD-256 as a colorless gel (61.2 mg 70%over 4 steps).

491-HAD-260

Reaction mixture containing 491-HAD-256 (59.1 mg, 0.0739 mmol),molecular sieves (4 47.8 mg), Celite (47.8 mg) and PCC (47.8 mg, 0.222mmol) in CH₂Cl₂ (1.70 mL) was stirred under a nitrogen atmosphere at rtfor 1 hr. Triethylamine (30.8 μL, 0.222 mmol) was added and reactionmixture was stirred for an additional 15 min. Diethyl ether was addedand reaction mixture was filtered through Celite. Volume of the filtratewas reduced and flash chromatography (100% EtOAc) afforded slightlyimpure 491-HAD-260 as a colorless oil (90.9 mg).

491-HAD-261

To a solution of 491-HAD-260 in CH₂Cl₂ (0.60 mL), under a nitrogenatmosphere, TFA was added. After stirring 30 min at rt, solvent andvolatiles were removed by rotary evaporation. Acetonitrile (0.6 mL) and1 N HCl (0.60 mL, 0.60 mmol) were added to the residue. After stirring19 h at rt, reaction mixture was cooled with ice/water bath and asaturated aqueous solution of NaHCO₃ was added. Aqueous phase wasextracted several times with CHCl₃ then once with 10% methanol/CHCl₃.Combined CHCl₃ extracts were dried with anhydrous Na₂SO₄ andconcentrated by reduced pressure. Flash chromatography (MeOH: CH₂Cl₂: 2MNH₃/MeOH 5:95:1, 10:90:1, 15:85:1) yielded 491-HAD-261, ER805023 as anoff-white solid (6.8 mg, 18% over 3 steps).

Preparation of Compound ER-804446 (C14 Difluoromethoxy)

Step 1

To a solution of compound 557-MS-262 (4.14 g, 6.69 mmol) in THF (40 mL)was added a 1M solution of TBAF (6.69 mL; 6.69 mmol). The reactionmixture was stirred at room temperature for 30 minutes then worked up inthe usual manner. Chromatographic purification gave compound 557-MS-151(2.34 g, 92%).

Step 2

To a vigorously stirred solution of compound 557-MS-151 (520 mg, 1.36mmol) in dioxane (2 mL) at 55-60° C., was added pre-heated (50° C.) 40%aqueous NaOH solution (2 mL). A stream of chlorodifluoromethane gas wasthen admitted continuously to the reaction mixture via a gas inlet tube(the tip of which was positioned just below the surface of the reactionmixture). After 25 minutes the reaction mixture was worked up in theusual manner. Chromatographic purification gave compound 557-MS-154 (329mg, 56%).

Step 3

A solution of 557-MS-154 (455 mg, 1.05 mmol) in ethanol (5 mL) wastreated with 40% aqueous NaOH solution (2 mL) and heated under refluxfor 16 hours. The reaction mixture was cooled to room temperature,diluted with water and then washed with diethyl ether. The aqueous phasewas acidified (with cooling) to pH3 by drop wise addition ofconcentrated aqueous hydrochloric acid. Extraction with diethyl ether(×4) followed by drying etc gave compound 557-MS-158 (384 mg, 88%).

Step 4

A solution of compound 557-MS-158 (384 mg, 0.92 mmol) in diethyl ether(8 mL) was treated with toluene (2 mL), triphenylphosphine (290 mg, 1.10mmol), and 2-(trimethylsilyl)ethanol (0.172 ml, 1.20 mmol), then cooledto 0° C. under an inert atmosphere. Diethyl azidodicarboxylate (0.174ml, 1.10 mmol) was added drop wise and the reaction mixture then allowedto warm to room temperature. After 3 hours the reaction mixture wasworked up in the usual manner. Chromatographic purification gavecompound 557-MS-165 (410 mg, 86%).

Step 5

A mixture of compound 557-MS-165 (410 mg, 0.79 mmol) and compound554-RB-260 (523 mg, 0.72 mmol) was dissolved in THF (3.2 mL), treatedwith HMPA (0.6 mL) and then cooled to −78° C. under an inert atmosphere.A 0.5M solution of LiHMDS in THF (1.73 mL, 0.864 mmol) was then addeddrop wise over approximately 15 minutes. The reaction mixture wasstirred at −78° C. for 40 minutes, then warmed to 0° C. The intermediatecrude product was worked up in the usual manner and then dissolved indichloromethane (12 mL) and cooled to 0° C. A solution of approximately55% m-CPBA (452 mg) in dichloromethane (8 mL) was added portion wise.After 40 minutes triethylamine (1 mL) was added and the reaction mixturewas worked up in the usual manner. Chromatographic purification gavecompound 557-MS-203 (552 mg, 80%).

Step 6

A solution of compound 557-MS-203 (552 mg, 0.575 mmol) in THF (5.75 mL)was treated with a 1M solution of TBAF in THF (11.5 mL, 11.5 mmol) thenheated at 60° C. for approximately 3 hours. The usual work up gave crudecompound 557-MS-205 (350 mg), which was used in the next stage withoutpurification.

Step 7

A solution of crude compound 557-MS-205 (assumed to contain 0.32 mmol)in dichloroethane (66 mL) was added slowly to a heated solution (85° C.)of 2-chloro-1-methylpyridinium iodide (824 mg, 3.2 mmol) andtri-n-butylamine (0.768 mL, 3.2 mmol) in dichloroethane (100 mL). Thereaction mixture was heated at 85° C. for 1 hour post complete additionthen cooled to room temperature. The usual work up and chromatographicpurification gave compound 557-MS-212 (93 mg, 48% from compound557-MS-203).

Step 8

A solution of compound 557-MS-212 (93 mg, 0.15 mmol) in THF (6 mL) andethanol (12 mL) was treated with 1M aqueous NaOH solution (2.5 mL) andheated at 60° C. for 1.5 hours, then at 70° C. for 1 hour. The usualwork up gave crude compound 557-MS-214 (74 mg), which was used in thenext stage without purification.

Step 9

A solution of crude compound 557-MS-214 (89 mg, 0.178 mmol) indichloromethane (10 mL) was treated with PCC (462 mg, 2.14 mmol) in thepresence of powdered 4 Å molecular sieves (462 mg). The reaction mixturewas stirred vigorously for 120 minutes at room temperature. Basificationwith excess triethylamine, followed by chromatographic purification gavecompound 557-MS-216 (53 mg, 60% from compound 557-MS-212).

Step 10

A solution of compound 557-MS-216 (53 mg, 0.106 mmol) in a mixture ofacetonitrile (7 mL) and dichloromethane (1.7 mL) was treated with 48%aqueous hydrofluoric acid (1.7 mL). After 35 minutes at room temperaturethe usual work up, followed by chromatographic purification, gavecompound ER-804446 (40 mg, 91%) (m/z: 411.3 [M+1; 100%]).

Preparation of Compound ER-804387 (C14 Trifluoroethoxy)

Step 1

To a solution of 557-MS-151 (1 g, 2.62 mmol) in acetone (20 mL) wereadded potassium carbonate (440 mg, 3.14 mmol) and 2,2,2-trifluoroethyltrichloromethanesulphonate (880 mg, 3.14 mmol). The reaction mixture washeated at 70° C. for 2 hours then treated with further aliquots ofpotassium carbonate (440 mg; 3.14 mmol) and 2,2,2-trifluoroethyltrichloromethanesulphonate (880 mg, 3.14 mmol). After a further 2 hoursat 70° C. the reaction mixture was worked up in the usual manner.Chromatographic purification gave compound 557-MS-161 (760 mg, 63%).

Step 2

A solution of 557-MS-161 (760 mg, 1.64 mmol) in ethanol (5 mL) wastreated with 40% aqueous NaOH solution (2 mL) and heated under refluxfor 16 hours. The reaction mixture was cooled to room temperature,diluted with water and then washed with diethyl ether. The aqueous phasewas acidified (with cooling) to pH3 by drop wise addition ofconcentrated aqueous hydrochloric acid. Extraction with diethyl ether(×4) followed by drying etc gave compound 557-MS-163 (648 mg, 88%).

Step 3

A solution of compound 557-MS-163 (648 mg, 1.44 mmol) in diethyl ether(12 mL) was treated with toluene (3 mL), triphenylphosphine (454 mg,1.73 mmol), and 2-(trimethylsilyl)ethanol (0.269 mL, 1.875 mmol), thencooled to 0° C. under an inert atmosphere. Diethyl azidodicarboxylate(0.272 mL, 1.73 mmol) was added drop wise and the reaction mixture thenallowed to be warmed to room temperature. After 1.5 hours the reactionmixture was worked up in the usual manner. Chromatographic purificationgave compound 557-MS-167 (730 mg, 92%).

Step 4

A mixture of compound 557-MS-167 (730 mg, 1.33 mmol) and compound554-RB-260 (885 mg, 1.21 mmol) was dissolved in THF (9 mL), treated withHMPA (1 mL) and then cooled to −78° C. under an inert atmosphere. A 0.5Msolution of LiHMDS in THF (2.9 mL, 1.45 mmol) was then added drop wiseover approximately 20 minutes. The reaction mixture was stirred at −78°C. for 35 minutes, then warmed to 0° C. The intermediate crude productwas worked up in the usual manner and purified partially bychromatography. The intermediate was dissolved in dichloromethane (15mL) and cooled to 0° C. A solution of approximately 55%meta-chloroperbenzoic acid (612 mg) in dichloromethane (10 mL) was addedportion wise. After 30 minutes triethylamine (1.37 mL) was added and theusual work up, followed by chromatographic partial purification, gaveimpure compound 557-MS-177 (950 mg), which was used in the next stagewithout further purification.

Step 5

A solution of crude compound 557-MS-177 (475 mg, assumed to contain0.479 mmol) in a 1M solution of TBAF in THF (9.58 mL; 9.58 mmol) washeated at 50° C. for approximately 7 hours. The usual work up gave crudecompound 557-MS-179 (300 mg), which was used in the next stage withoutpurification.

Step 6

A solution of crude compound 557-MS-179 (assumed to contain 0.153 mmol)in dichloroethane (30 mL) was added slowly to a heated solution (85° C.)of 2-chloro-1-methylpyridinium iodide (391 mg, 1.53 mmol) andtri-n-butylamine (0.365 mL, 1.53 mmol) in dichloroethane (100 mL). Thereaction mixture was heated at 85° C. for 1 hour post complete additionthen cooled to room temperature. The usual work up and chromatographicpurification gave compound 557-MS-183 (40 mg, 41% from compound557-MS-167).

Step 7

A solution of compound 557-MS-183 (40 mg, 0.063 mmol) in THF (2.5 mL)and ethanol (5 mL) was treated with 1M aqueous NaOH solution (1 mL) andheated at 60° C. for 3.5 hours. The usual work up gave crude compound557-MS-191 (32 mg), which was used in the next stage withoutpurification.

Step 8

A solution of crude compound 557-MS-191 (52 mg, 0.098 mmol) indichloromethane (6 mL) was treated with PCC (253 mg, 1.18 mmol) in thepresence of powdered 4 Å molecular sieves (253 mg). The reaction mixturewas stirred vigorously for 3 hours at room temperature. Basificationwith excess triethylamine, followed by chromatographic purification gavecompound 557-MS-194 (39.3 mg, 76% from compound 557-MS-183).

Step 9

A solution of compound 557-MS-194 (23 mg, 0.0435 mmol) in a mixture ofacetonitrile (3.2 mL) and dichloromethane (0.8 mL) was treated with 48%aqueous hydrofluoric acid (0.8 mL). After 35 minutes at room temperaturethe usual work up, followed by chromatographic purification, gavecompound ER-804387 (15.8 mg, 82%).

Preparation of Compound B2356 (C14 Hydroxy) & Compound B2359 (C14 OMOM):

Step 1

A solution of methyl 2,4-dihydroxy-6-methylbenzoate (6 g, 32.95 mmol) indry DMF (10 mL) was added to a well-stirred suspension of hexane-washedsodium hydride (3.95 g, 60% in mineral oil; approximately 98.85 mmol) indry DMF (50 mL), at 0° C. under an inert atmosphere. The reactionmixture was stirred at 0° C. for 30 minutes then treated withmethoxymethyl chloride (5.26 mL, 69.2 mmol) drop wise. The reactionmixture was stiffed for 2 hours then worked up in the usual manner togive compound 453-MS-21 (8.14 g, 91%).

Step 2

To a solution of diisopropylamine (3.14 mL, 22.4 mmol) in dry THF (45mL), at −20° C. under an inert atmosphere, was added drop wise a 2.5Msolution of n-butyllithium in hexanes (8.96 mL, 22.4 mmol). The reactionmixture was allowed to warm to 0° C., stirred for 10 minutes at 0° C.,then cooled to −78° C. A solution of compound 453-MS-21 (4.03 g, 14.9mmol) in dry THF (15 mL) was added drop wise. The reaction mixture wasstirred at −78° C. for 1 hour then treated with a solution of diphenyldiselenide (5.59 g, 17.9 mmol) in dry THF (18 mL). The reaction mixturewas stirred at −78° C. for 30 minutes then worked up in the usualmanner. Chromatographic purification gave compound 453-MS-108 (3.47 g,54%).

Step 3

A solution of compound 453-MS-108 (2.16 g, 5.08 mmol) in ethanol (20 mL)was treated with powdered NaOH (610 mg, 15.24 mmol) and heated underreflux for 28 hours. The reaction mixture was cooled to room temperatureand concentrated under reduced pressure (to a residual volume ofapproximately 5 mL). The residue was partitioned between water anddiethyl ether. The aqueous fraction was acidified to pH3 by slowaddition of 1M aqueous HCl (approximately 16 mL), with cooling. Theacidic solution was extracted with diethyl ether, and the extracts thenwashed immediately with saturated aqueous brine solution (at least fivetimes). Drying etc gave compound 453-MS-110 (2.016 g, 97%).

Step 4

To a solution of compound 453-MS-110 (2.016 g, 4.9 mmol) in diethylether (40 mL), at 0° C. under an inert atmosphere, were added toluene(10 mL), triphenylphosphine (1.41 g, 5.39 mmol), and2-(trimethylsilyl)ethanol (0.843 mL, 5.88 mmol). Diethylazidodicarboxylate (0.849 mL, 5.39 mmol) was then added drop wise. Thereaction mixture was allowed to warm to room temperature and stirred forapproximately 16 hours. The usual work up followed by chromatographicpurification gave compound 453-MS-111 (2.24 g, 90%).

Step 5

A mixture of compound 453-MS-111 (1.33 g, 2.6 mmol) and compound343-YW-281 (866 mg, 1.46 mmol) was dissolved in a solution of 10% HMPAin THF (15 mL) and cooled to −78° C. under an inert atmosphere. A 1Msolution of LiHMDS in THF (2.19 mL, 2.19 mmol) was then added drop wiseover approximately 15 minutes. After another 45 minutes an extra aliquotof 1M solution of LiHMDS in THF was added (0.438 mL, 0.438 mmol). Thereaction mixture was warmed to 0° C. and the intermediate crude productworked up in the usual manner, and purified partially by chromatography.The intermediate was then dissolved in dichloromethane (14 mL) andcooled to 0° C. A solution of approximately 55% meta-chloroperbenzoicacid (280 mg) in dichloromethane (6 mL) was added. After 10 minutes,extra 55% m-CPBA (28 mg) was added. The reaction mixture was stirred at0° C. for a further 20 minutes, then treated with triethylamine (1.22mL) and worked up in the usual manner. Chromatographic purification gavecompound 453-MS-77 (625 mg, 52%).

Step 6

To a vigorously stirred biphasic mixture of compound 453-MS-77 (618 mg,0.753 mmol), dichloromethane (20 mL) and water (10 mL), was added DDQ(190 mg, 0.84 mmol). After 1 hour at room temperature the reactionmixture was worked up in the usual manner. Chromatographic purificationgave compound 453-MS-82 as a mixture of 4 diastereoisomers (381 mg,72%).

Step 7

A solution of compound 453-MS-82 (381 mg, 0.544 mmol) in THF (10 mL) wastreated with TBAF (284 mg, 1.09 mmol) and stirred at room temperaturefor approximately 16 hours. The usual work up gave compound 453-MS-84(326 mg, quantitative), as a mixture of 4 diastereoisomers.

Step 8

To a solution of triphenylphosphine (109 mg, 0.417 mmol) in dry THF (6mL), at room temperature under an inert atmosphere, was added diethylazidodicarboxylate (66 μL, 0.417 mmol) drop wise over approximately 30seconds. A solution of compound 453-MS-84 (167 mg, 0.278 mmol) in dryTHF (10 mL) was added drop wise over approximately 10 minutes. Thereaction mixture was stirred at room temperature for 10 minutes thentreated with extra triphenylphosphine (36 mg, 0.137 mmol), followed byextra diethyl azidodicarboxylate (22 μL, 0.137 mmol). The reactionmixture was stirred for a further 10 minutes then worked up in the usualmanner. Partial chromatographic purification gave compound 453-MS-91(122 mg, 76%) as a mixture of 4 diastereoisomers.

Step 9

A solution of compound 453-MS-91 (70 mg, 0.12 mmol) in ethanol (2.5 mL)was treated with THF (1.25 mL) and 1M aqueous NaOH (0.6 mL, 0.6 mmol)and stirred at room temperature for approximately 6 days.Chromatographic purification gave two fractions of partially resolveddiastereoisomers:

Fraction A (less polar): a mixture of 2 diastereoisomers—compound453-MS-101A (24 mg);

Fraction B (more polar): a mixture of 2 diastereoisomers—compound453-MS-101B (25 mg);

(total yield: 49 mg, 86%)

Step 10

A solution of compound 453-MS-101B (25 mg, 52.3 μmol) in dichloromethane(1.5 mL) was treated with PCC (135 mg, 0.627 mmol) in the presence ofpowdered 4 Å molecular sieves (135 mg). The reaction mixture was stirredvigorously for 40 minutes at mom temperature. Basification with excesstriethylamine, followed by chromatographic purification gave compound453-MS-116 (14 mg, 63%).

Step 11

A solution of compound 453-MS-116 (13 mg, 27.3 μmol) in a mixture ofdioxane (3 mL) and deuterium oxide (3 mL) was treated with Dowex®(50WX8-100, 200 mg), and stirred at room temperature for approximately16 hours. The usual work up, followed by purification using reversedphase HPLC, gave compound B2356 (2.4 mg, 25%) [m/z: 349.3 (M+1, 60%),161.0 (100%)], and compound B2358 (2 mg, 20%).

Preparation of Compound B2357 (C14 Hydroxy) & Compound B2359 (C14 OMOM):

Step 1

A solution of compound 453-MS-101A (24 mg, 50.1 μmol) in dichloromethane(2 mL) was treated with pyridinium chlorochromate (130 mg, 0.602 mmol)in the presence of powdered 4 Å molecular sieves (130 mg). The reactionmixture was stirred vigorously for 1 hour at room temperature.Basification with excess triethylamine, followed by chromatographicpurification gave compound 453-MS-122 (19 mg, 80%).

Step 2

A solution of compound 453-MS-122 (21 mg, 44 μmol) in a mixture ofacetonitrile (4.6 mL) and dichloromethane (1.1 mL) was treated with 48%aqueous hydrofluoric acid (1.1 mL), and stirred at room temperature forapproximately 2 hours. The usual work up, followed by purification usingreversed phase HPLC, gave compound B2357 (3.5 mg, 23%) [m/z: 349.2 (M+1,50%), 161.0 (100%)], and compound B2359 (1.6 mg, 10%).

Preparation of C14-C, H, or Halogen Analogs, NF0887, NF2433, NF2435,NF2436, NF2557 and ER-805053,

Synthetic Procedure for NF2433

Using same procedure for 16, YE-06 (526 mg, 0.714 mmol) was converted toNY-78 (657 mg).

Using same procedure for 18, NY-78 (653 mg, 0.644 mmol) was converted toNY-79 (529 mg).

Using same procedure for 509-HD-116, NY-79 (528 mg, 0.584 mmol) wasconverted to NY-80 (231 mg). NY-80 was used without purification for thenext step.

Using same procedure for TM-12, NY-80 (230 mg, 0.511 mmol) was convertedto NY-81 (157 mg).

Using same procedure for 509-HD-125, NY-81 (127 mg, 0.294 mmol) wasconverted to NY-82 (118 mg).

Using same procedure for NF-0675, NY-82 (118 mg, 0.274 mmol) wasconverted to NF-2433 (79 mg).

Synthetic Procedure for NF-2436

Using same procedure for 9, NY-83 (7.46 g, 40 mmol) was converted toNY-84 (11.39 g).

To a mixture of NY-84 (3 g, 10 mmol), Et₂NH (2.07 mL, 20 mmol) and THF(80 mL), isopropylmagenesium chloride (2M in THF, 10 mL, 20 mmol) wasgradually added at −30° C. The reaction mixture was allowed to warm to0° C. The reaction mixture was quenched with sat. NaHCO₃ and extractedwith EtOAc. The organic layer was washed with water, brine, dried overNa₂SO₄, filtered and concentrated. The crude product was purified onsilica gel column with hexane/EtOAc, 10:1, 8:1, 6:1 to give 3.21 g ofNY-85.

NY-85 (3.2 g, 9.36 mmol) and TMEDA (2.2 mL, 14.58 mmol) were dissolvedin THF (30 mL) and cooled to −78° C., under nitrogen. Then, sec-BuLi(1.3M/cyclohexane, 11 mL, 14.3 mmol) was slowly added and the reactionwas stirred at −78° C. for 1 hr. DMF (1.4 mL, 18.08 mmol) was added tothe solution, then the solution was stirred at −78° C. for 30 min. Themixture was quenched with sat. NH₄Cl and extracted with EtOAc. Theorganic layer was washed with water, brine and dried over Na₂SO₄,filtered and concentrated. The crude product was purified on silica gelcolumn with hexane/EtOAc, 6:1 to give 2.56 g of NY-86.

To a solution of NY-86 (2.55 g, 6.89 mmol) in MeOH (30 mL), NaBH₄ (260mg, 6.87 mmol) was added at 0° C. and stirred for 30 min. The reactionmixture was quenched with sat. NH₄Cl and extracted with EtOAc. Theorganic layer was washed with water, brine, dried over Na₂SO₄, filteredand concentrated to give 2.5 g of NY-87.

A mixture of NY-87 (2.5 g, 6.72 mmol), AcOH (1.92 mL, 33.54 mmol) andEtOH (50 mL) was refluxed for 4 hrs. The mixture was concentrated andthe residue was alkalized with sat. NaHCO₃ and extracted with EtOAc. Theaqueous layer was reextracted with EtOAc (×2) and the organic layerswere washed with sat. NH₄Cl, water, brine, dried over Na₂SO₄, filteredand concentrated. The crude crystallized product was washed with Et₂O togive 718 mg of NY-88. The mother liquid was purified on silica gelcolumn with hexane/EtOAc, 3:1, 2:1 to give additional 230 mg of NY-88.

Using same procedure for 509-HD-209, NY-88 (940 mg, 5.09 mmol) wasconverted to NY-89 (676 mg).

To a solution of NY-89 (1.74 g, 7.61 mmol) in DMSO (20 mL), a solutionof KOH (450 mg, 8.02 mmol) in water (10 mL) was added and stirred atroom temperature for 30 min. Then, the mixture was stirred at 40° C. for30 min. MeI (9.5 mL, 153 mmol) was added to the reaction mixture, andthe mixture was stirred at room temperature for 3 hrs. The reactionmixture was quenched with sat. NH₄Cl and extracted with EtOAc. Theorganic layer was washed with water, 5% citric acid, water, sat. NaHCO₃,brine, dried over Na₂SO₄, filtered and concentrated to give 1.93 g ofNY-90.

To a solution of NY-90 (1.93 g, 7.61 mmol), Ph₂S₂ (5 g, 22.9 mmol),pyridine (3.7 mL, 45.75 mmol), ^(n)Bu₃P (5.7 mL, 22.88 mmol) were addedat 0° C. The mixture was allowed to warm to room temperature and stirredfor 12 hrs. The reaction mixture was diluted with EtOAc and washed with5% citric acid (×2), water, sat. NaHCO₃, brine, dried over Na₂SO₄,filtered and concentrated. The crude product was purified on silica gelcolumn with hexane/EtOAc, 15:1, 5:1 to give 1.9 g of NY-91.

Using same procedure for 509-HD-212, NY-91 (2.12 g, 6 mmol) wasconverted to NY-92 (2.07 g). NY-92 was used without purification for thenext step.

Using same procedure for 509-HD-213, NY-92 (2.07 g, 6 mmol) wasconverted to NY-93 (2.19 g).

Using same procedure for 16, YE-06 (505 mg, 0.685 mmol) was converted toNY-94 (625 mg).

Using same procedure for 18, NY-94 (623 mg, 0.594 mmol) was converted toNY-95 (501 mg).

Using same procedure for 509-HD-116, NY-95 (500 mg, 0.533 mmol) wasconverted to NY-96 (400 mg). NY-96 was used without purification for thenext step.

Using same procedure for TM-12, NY-96 (400 mg, 0.533 mmol) was convertedto NY-97 (142 mg).

Using same procedure for 509-HD-125, NY-97 (139 mg, 0.298 mmol) wasconverted to NY-98 (120 mg).

Using same procedure for NF-0675, NY-98 (118 mg, 0.254 mmol) wasconverted to NF-2436 (24 mg).

Synthetic Procedure for NF2435

Using similar procedure for the synthesis of 509-HD-209 from 509-HD-207,3,5-dimethylphenol (27.58 g, 225.75 mmol) was converted to colorless oilof MK-091 (15.91 g, 42%) as compound purified.

MK-091 (6.00 g, 36.10 mmol) was dissolved in 18 mL of dry Et₂O. To thestirred solution was added n-BuLi in hexane (1.6M, 27 mL, 43.32 mmol,1.2 eq.) and the mixture was warmed to 40° C. After 30 min at 40° C. thecolor of the reaction mixture turned dark red. The mixture was cooled to−78° C. and excessive dry CO₂ gas (ca 30 eq.) was added by bubblingthrough an inlet over 30 min. Then, the resulting mixture was allowed towarm to rt. After 30 min the reaction mixture was quenched with waterand washed with Et₂O. The basic aqueous layer was acidified with aqueoussolution of KHSO₄ and extracted with AcOEt. The organic extract waswashed with brine, dried over Na₂SO₄, filtered, and concentrated toyield crude colorless crystals of MK-092 (4.55 g, <60%). The crudeMK-092 was used for next step without purification.

Crude MK-092 (4.55 g, assumed to contain 21.6 mmol) was dissolved in 200mL of CH₃CN. To the solution were added CsCO₃ (5.64 g, 17.3 mmol) andMeI (2.20 mL, 34.6 mmol) and the mixture was stirred at rt overnight.After concentration of the reaction mixture, water was added andextracted with AcOEt. The organic extract was washed with saturatedaqueous NaHCO₃ and brine, then dried over Na₂SO₄, filtered, andconcentrated to give crude oil of MK-093. The crude product was purifiedby chromatography on silica gel (hexane/AcOEt: 10/1) to afford paleyellow oil of MK-093 (3.19 g, 39% 2 steps).

Using similar procedure for the synthesis of 509-HD-211 from 509-HD-209,MK-093 (2.86 g, 12.77 mmol) was converted to a mixture of MK-094, MK-095and MK-093. The crude product was purified by chromatography on silicagel (hexane/AcOEt: 9/1) to afford pale yellow oil of MK-094 (652 mg,13%), the structure of which was determined by NOESY analysis.

Using similar procedure for the synthesis of 509-HD-212 from 509-HD-211,MK-094 (650 mg, 1.71 mmol) was converted to crude benzoic acid.

Using similar procedure for the synthesis of MK-047 from MK-046, thecrude benzoic acid was converted to pale pink oil of MK-096 (746 mg, 94%2 steps) as compound purified.

Using similar procedure for the synthesis of compound 5 from compound 2and compound 3, YE-06 (516 mg, 0.700 mmol) coupled with MK-096 (489 mg,1.05 mmol) was converted to colorless oil of MK-097 (491 mg, 76% 3steps) as compound purified.

Using similar procedure for the synthesis of YE-17 from YE-16, MK-097(491 mg, 0.535 mmol) was converted to crude oil of MK-098 (486 mg,including silyl impurity). The crude MK-098 was used for next stepwithout purification.

Using similar procedure for the synthesis of YE-18 from YE-17, the crudeMK-098 (486 mg, assumed to contain 0.535 mmol) was converted tocolorless oil of MK-099 (159 mg, 67% 3 steps) as compound purified.

Using similar procedure for the synthesis of 509-HD-125 from509-HD-119B, MK-099 (158 mg, 0.354 mmol) was converted to crude paleyellow solid of MK-100 (146 mg, <93%). The crude MK-100 was used fornext step without purification.

Using similar procedure for the synthesis of NF1226 from MK-035, crudeMK-100 (146 mg) was converted to crude pale yellow crystals. The crudeproduct was purified by chromatography on silica gel (hexane/AcOEt: 3/2)to afford colorless crystals of NF2435 (88 mg, 69% 2 steps).

Synthetic Procedure for NF2557, NF2558, and NF2559

Using similar procedure for the synthesis of 509-HD-209 from 509-HD-207,TM-34 (12.23 g, 42.71 mmol) was converted to colorless oil of MK-120(13.40 g, 95%) as compound purified.

MK-120 (13.38 g, 40.50 mmol) was dissolved in 200 mL of EtOH. 10% Pd oncarbon (50% wet, 2.7 g) was added. The mixture was stirred underhydrogen at rt. After 8 hrs catalyst was filtered through Celite and thefiltrate was concentrated to give crude solid. The crude product waspurified by chromatography on silica gel (hexane/AcOEt: 2/1) to affordcolorless crystals of MK-121 (8.67 g, 89%).

Using similar procedure for the synthesis of 611-MS-88 from 611-MS-84,MK-121 (8.66 g, 36.04 mmol) was converted to colorless crystals ofMK-122 (13.20 g, 98%) as compound purified.

Using similar procedure for the synthesis of MK-073 from MK-072,propargyl alcohol (10.74 g, 191.6 mmol) was converted to colorless oilof MK-123 (27.03 g, 96%) as compound purified.

MK-122 (13.20 g, 35.46 mmol) and MK-123 (12.50 g, 70.92 mmol, 2.0 eq.)were dissolved in 440 mL of DMF. To the solution were added Ph₃P (5.58g, 21.28 mmol, 0.6 eq.), Pd(Ph₃P)₄ (6.15 g, 5.32 mmol, 0.15 eq.), CuI(1.01 g, 5.32 mmol, 0.15 eq.), and Et₃N (19.8 mL, 141.83 mmol, 4 eq.).The mixture was heated to 45° C. and stirred for 1.5 hrs under nitrogenatmosphere. The mixture was cooled to rt and diluted with Et₂O-hexane,then stirred for a while. The resulting mixture was filtered through apad of Celite to remove insoluble solid. The filtrate was washed withsaturated aqueous solution of NH₄Cl and brine, dried over Na₂SO₄,filtered, and concentrated to yield crude product. The crude product waspurified by chromatography on silica gel (hexane/AcOEt: 7/1 to 5/1) toafford pale brown oil of MK-124 (12.11 g, 86%).

Using similar procedure for the synthesis of MK-074 from MK-073, MK-124(14.28 g, 35.84 mmol) was converted to crude yellow oil of MK-125 (14.34g). The crude MK-125 was used for next step without purification.

Using similar procedure for the synthesis of MK-114 from MK-113, crudeMK-125 (14.34 g, assumed to contain 35.84 mmol) was converted to crudeyellow oil of benzoic acid (14.92 g). The crude benzoic acid was usedfor next step without purification.

Using similar procedure for the synthesis of MK-093 from MK-092, thecrude benzoic acid (14.92 g, assumed to contain 35.84 mmol) wasconverted to colorless oil of MK-126 (10.92 g, 78% 3 steps) as compoundpurified.

Using modified procedure for the synthesis of 509-HD-211 from509-HD-209, MK-126 (960 mg, 2.47 mmol) was converted to crude selenideproduct. The crude product was purified by chromatography on silica gel(hexane/AcOEt: 6/1 to 5/1) to afford pale yellow oil (953 mg) as aninseparable mixture of desirable MK-127 (ca 710 mg, ca 60%), MK-128 (ca10%), and MK-126 (ca 17%). The mixture was used for next step withoutfurther purification.

Using similar procedure for the synthesis of MK-114 from MK-113, themixture (953 mg) of MK-127 (ca 710 mg, assumed to contain 1.31 mmol),MK-128, and MK-126 was converted to crude oil of benzoic acids (980 mg).The crude benzoic acids were used for next step without purification.

Using similar procedure for the synthesis of MK-047 from MK-046, thecrude mixture of benzoic acids (980 mg) was converted to crude TMS-ethylesters. The crude product was purified by chromatography on silica gel(hexane/AcOEt: 6/1) to afford colorless oil (875 mg) as an inseparablemixture of desirable MK-129 (ca 683 mg, ca 83%) and other TMS-ethylesters corresponding to MK-128 and MK-126. The mixture was used for nextstep without further purification.

Using similar procedure for the synthesis of compound 5 from compound 2and compound 3, YE-06 (483 mg, 0.655 mmol) coupled with the mixture (875mg) including MK-129 (ca 683 mg, 1.08 mmol) was converted to colorlessoil of MK-130 (519 mg, 73% 3 steps).

Using similar procedure for the synthesis of YE-17 from YE-16, MK-130(519 mg, 0.480 mmol) was converted to crude oil of MK-131 (620 mg,including silyl impurity). The crude product was used for next stepwithout purification.

Using similar procedure for the synthesis of YE-18 from YE-17, the crudeMK-131 (620 mg, assumed to contain 0.480 mmol) was converted to crudeoil of lactonized product. The crude product was purified bychromatography on silica gel (hexane/AcOEt: 5/2) to afford colorless oilof MK-132 (88 mg, 30% 3 steps) and colorless oil of des-MOM form MK-133(47 mg, 17% 3 steps) respectively.

Using similar procedure for the synthesis of 509-HD-125 from509-HD-119B, MK-133 (46 mg, 0.0812 mmol) was converted to crude paleyellow oil of enone (35 mg, <76%). The crude enone was used for nextstep without purification.

Using similar procedure for the synthesis of NF0531 from NF0530, thecrude enone (35 mg, assumed to contain 0.0620 mmol) was converted tocolorless crystals of MK-134 (20 mg, 55% 2 steps).

Using similar procedure for the synthesis of NF0675 from TM-13, MK-134(7 mg, 0.0157 mmol) was converted to colorless crystals of NF2557 (5.1mg, 80%) and colorless solid of NF2558 (1.5 mg, 19%) as compoundpurified, respectively.

Synthesis of ER-805053

Step 1

To a solution of methyl 2,4-dihydroxy-6-methylbenzoate (10.9 g, 0.0598mol) in dry N,N-dimethylformamide (100 mL) were added imidazole (4.48 g,0.0658 mol) and tert-butyldiphenylsilyl chloride (15.6 mL, 0.0658 mol).The reaction mixture was stirred at room temperature for 24 hours thenworked up in the usual manner. The crude product was purifiedchromatographically to give compound 557-MS-232 (12.33 g, 49%).

Step 2

To a solution of compound 557-MS-232 (9.08 g, 0.021 mol) in drytetrahydrofuran (100 mL), at 0° C. under an inert atmosphere, was addedsodium hydride (55% dispersion in oil; 1.88 g, approximately 0.042 mol).The reaction mixture was stirred at 0° C. for 30 minutes then treatedwith methoxymethyl chloride (3.28 mL, 0.042 mol). The reaction mixturewas allowed to warm to room temperature overnight. The usual work upgave compound 557-MS-233 (9.17 g, 91%).

Step 3

To a solution of freshly prepared lithium diisopropylamide (16.2 mmol)in dry tetrahydrofuran (15 mL), at −78° C. under an inert atmosphere,was added drop wise a solution of compound 557-MS-233 (3.93 g, 9.01mmol) in dry tetrahydrofuran (15 mL). The reaction mixture was stirredat −78° C. for 45 minutes then a solution of diphenyl diselenide (2.53g, 8.11 mmol) in dry tetrahydrofuran (15 mL) was added rapidly (down theinside walls of the reaction vessel so as to pre-chill the diphenyldiselenide solution). The reaction mixture was stirred at −78° C. for 45minutes then treated drop wise with a 2M solution of acetic acid indiethyl ether (8 mL). The reaction mixture was then worked up in theusual manner. Chromatographic purification gave compound 557-MS-262(4.57 g, 82%).

Step 4

A solution of compound 557-MS-262 (6.29 g, 0.01 mol) in ethanol (100 mL)was treated with powdered sodium hydroxide (4 g, 0.1 mol) and heatedunder reflux for 30 minutes. The reaction mixture was cooled to 5° C.and acidified to pH6.5 with 1M HCl. The majority of the ethanol was thenremoved by concentration in vacuo and resultant residue partitionedbetween water and ethyl acetate. The organic layer was washedsequentially with saturated aqueous sodium bicarbonate solution andwater. Drying etc gave a crude residue, which was purifiedchromatographically to give compound 611-MS-84 (3.3 g, 87%).

Step 5

A solution of compound 611-MS-84 (1.65 g, 4.33 mmol) in dichloromethane(20 mL), at 0° C., was treated sequentially with pyridine (0.385 mL,4.76 mmol) and trimethylsulfonic anhydride (0.764 mL, 4.54 mmol). After25 minutes at 0° C. the reaction mixture was warmed to room temperatureand worked up in the usual manner to give compound 611-MS-88 (1.5 g,68%).

Step 6

A solution of compound 611-MS-88 (1.5 g, 2.92 mmol) in1,2-dimethoxyethane (25 mL) was heated under reflux, under an inertatmosphere, in the presence of phenyl boronic acid (713 mg, 5.84 mmol),palladium tetrakistriphenylphosphine (335 mg, 0.29 mmol), lithiumchloride (247 mg, 5.84 mmol) and 2M aqueous sodium carbonate (25 mL).After 2 hours the reaction mixture was cooled to room temperature. Theusual work up, followed by chromatographic purification gave compound611-MS-91 (1.004 g, 78%).

Step 7

A mixture of compound 611-MS-91 (512 mg, 1.16 mmol) and compound554-RB-260 (635 mg, 1.05 mmol) was dissolved in a solution of 10%hexamethylphosphoramide in tetrahydrofuran (8.8 mL) and cooled to −78°C. under an inert atmosphere. A 0.5M solution of lithiumbis-(trimethylsilyl)amide in tetrahydrofuran (2.52 mL, 1.26 mmol) wasthen added drop wise over approximately 30 minutes. The reaction mixturewas stirred at −78° C. for 2 hours, then warmed to 0° C. Theintermediate crude product was worked up in the usual manner and thendissolved in dichloromethane (15 mL) and cooled to 0° C. A solution ofapproximately 55% meta-chloroperbenzoic acid (724 mg) in dichloromethane(12 mL) was added. After 30 minutes triethylamine (1.6 mL) was added andthe reaction mixture was worked up in the usual manner. Chromatographicpurification gave compound 611-MS-102 (560 mg, 64%).

Step 8

A solution of compound 611-MS-102 (560 mg, 0.738 mmol) intetrahydrofuran (5 mL) was treated with a 1M solution oftetrabutylammonium fluoride in tetrahydrofuran (0.74 mL, 0.74 mmol). Theusual work up, followed by chromatographic purification gave compound611-MS-104 (340 mg, 71%).

Step 9

A solution of compound 611-MS-104 (340 mg, 0.527 mmol) in ethanol (10mL) was treated with powdered sodium hydroxide (211 mg, 5.27 mmol) andheated under reflux. Cooling and acidification to pH 6.5, followed theusual work up gave crude compound 611-MS-106, which was used in the nextstage without purification.

Step 10

A solution of crude compound 611-MS-106 (assumed to contain 0.176 mmol)in dichloromethane (20 mL) was added slowly to a heated solution (40°C.) of 2-chloro-1-methylpyridinium iodide (449 mg, 1.76 mmol) andtri-n-butylamine (0.42 mL, 1.76 mmol) in dichloromethane (60 mL). Theusual work up and chromatographic purification gave compound 611-MS-108(3 mg, 3% from compound 611-MS-104).

Step 11

A solution of compound 611-MS-108 (3 mg, 5.9 μmol) in dichloromethane(500 μL) was treated with pyridinium chlorochromate (20 mg, 88 μmol) inthe presence of powdered 4 Å molecular sieves (20 mg). The reactionmixture was stirred vigorously for 4 hours at room temperature.Basification with excess triethylamine, followed by chromatographicpurification gave compound 611-MS-118 (1.4 mg, 48%).

Step 12

A solution of compound 611-MS-118 (1.4 mg, 2.76 μmol) in a mixture ofacetonitrile (400 μL) and dichloromethane (100 μL) was treated with 48%aqueous hydrofluoric acid (100 μL), and stirred at room temperature for30 minutes. The usual work up followed by chromatographic purificationgave compound ER-805053 (1.0 mg; 83%).

Preparation of C14-Aniline Analogs: ER805940, and ER806201:

Preparation of ER805940:

Tf₂O (0.42 ML, 2.5 mmole) was added to a solution of ER-805102 (0.95 g,1.7 mmole) and Et₃N (0.58 ML, 4.2 mmole) in 20 ML of CH₂Cl₂ at 0° C. Themixture was stirred for 10 min before the addition of aq. NaHCO₃. Aq.layer was extracted twice with CH₂Cl₂. The organics were concentratedand passed through a short plug of silica gel (20% EtOAc/Hex).

The triflate thus obtained was added Pd(OAc)₂ (19 mg, 0.08 mmole), BINAP(64 mg, 0.10 mmole) and Cs₂CO₃ (0.66 g, 2.0 mmole) in dry box.Benzophenone imine (0.32 ML, 1.9 mmole) and 30 ML of toluene was addedunder nitrogen before the mixture was heated at 90° C. for 14 h. Then itwas diluted with EtOAc and brine. Organic layer was dried (Na₂SO₄) andconcentrated.

Crude material was dissovled in 8 ML of MeOH and 5 ML of THF before theaddition of NaOAc (0.56 g, 6.8 mmole) and NH₂OH.HCl (0.24 g, 3.4 mmole)at RT. After 50 min, EtOAc and brine was added. Organics were dried(Na₂SO₄), concentrated and purified with silica gel (30 EtOAc/Hex) toproduce crystalline 629-ys-190 (0.88 g, 1.6 mmole).

LiHMDS (1N in THF, 8.0 mmole) was added slowly to a solution of629-ys-190 (0.88 g, 1.6 mmole) in 16 ML of THF at −55 to −50° C. Themixture was stirred at −45° C. for 5 min before the addition of BOC₂O(0.38 ML, 1.8 mmole). After the mixture was stirred at −40° C. for 30min, MeI (0.60 ML, 9.6 mmole) was added. After 10 min the mixture waswarmed up to RT for 2 h. Recooled to −35° C.,

the solution was added 72 ML of 1N NaOH and 48 ML of EtOH. After it washeated at 45° C. for 12 h, the mixture was diluted with 100 ML of waterand 150 ML of CH₂Cl₂. Aq. layer was extracted twice with 50 ML ofCH₂Cl₂. Organics were concentrated and purified by silica gelchromatography (30% EtOAc/Hex) to furnish colorless gel 629-ys-192 (0.58g, 1.0 mmole).

The suspension of 629-ys-192 (0.40 g, 0.71 mmole), PCC (0.46 g, 2.1mmole), 4 Å molecular sieves (0.50 g), and celite (0.50 g) in 8 ML ofCH₂Cl₂ was stirred at RT for 2.5 h before the addition of Et₃N (0.29 ML,2.1 mmole). After 5 min, 30 ML of Et₂O was added and the mixture wasfiltered. The filtrate was concentrated and passed through a shortsilica gel plug (75% EtOAc/Hex) to provide colorless crystalline629-ys-198 (0.35 g, 0.63 mmole).

TFA (5% water, 6 ML) was added slowly to the solution of 629-ys-198(0.35 g, 0.63 mmole) in CH₂Cl₂ at −35° C. The mixture was stirred at−20° C. for 1 h before the addition of sat. aq. NaHCO₃ (PH˜8) andCH₂Cl₂. Aq. layer was extracted twice with CH₂Cl₂. The organics weredried (Na₂SO₄), concentrated and purified by silica gel chromatography(75% EtOAc/Hex) to afford ER-805940 (124 mg, 0.33 mmole) in 25% overallyield over 8 steps.

Synthesis of ER806201:

1) Synthesis of Triflate:

To a solution of trihydroxy-benzoic acid (120 g) in 350 mL of acetone,500 mL of TFA (tri-fluoro acetic acid) was added at 40° C. understirring. After 1 h at that temperature, 300 mL of TFAA (tri-fluoroacetic anhydride) was added. The mixture was heated for 3 days. Themixture was distilled under house vacuum at 50° C. to remove solvents.The crude product was then diluted with 4 L of CH₂Cl₂, washed withwater, sat. NaHCO₃, dried and concentrated to give 85 g of semi puresolid. The solid was crystallized in EtOH (1 g/2 mL) to give 20 g ofpure crystal. The mother liq. was then purified by silica gel withCH₂Cl₂ to 5% MeOH/CH₂Cl₂ to give 55 g of additional product, 531-YW-184.

To a solution of 531-YW-184 (50 g, 238 mmol) in 156 mL of pyridine, Tf₂O(100 mL, 595 mmol, 2.5 eq.) was added at 0° C. in 3 h. Then it waswarmed to rt and stirred for 2 h. The reaction mixture was diluted withwater. The mixture was filtered. The solid on the filter was washed withwater, dried under vac to give a solid, 531-YW-187 (100 g).

To a mixture of ditriflate, 531-YW-187 (45.35 g), BocNH₂ (17.22 g),Pd₂(dba)₃ (4.38 g) and Pt-Bu₃ (4.38 g) in 150 mL of toluene,tri-ethylamine (26.92 mL) was added. The reaction was heated under inertatmosphere at 80° C. for 4 h. The crude reaction mixture was cooled andfiltered through a pad of celite. The filtrates were concentrated andpurified on silica gel with Hex/EtOAc, 9:1, 4:1 to give 28.3 g ofdesired product, 531-YW-194.

2) Synthesis of Olefin:

792-ANDI-114A was prepared analogously to the preparation of 554-RB-240with appropriate protecting groups, i.e. the MPM ether was replaced withTBDPS ether.

To a solution of 792-ANDI-114A (165.9 g, 265 mmol) in 2.65 L of Hexanes,quinoline (2.65 mL) and Lindlar catalyst (28.2 g, 13.3 mmol, 0.05 eq.)were added. The mixture was degassed repeatedly under vacuum andrecharged with nitrogen (3×) and hydrogen (3×). then it was set theintake of hydrogen on hydrogenator to 0.114 mol. The reaction wasmonitored by MS/1H NMR. After overnight, the suspension was filtered andrecharged with catalyst and hydrogen. After 3 days, the reaction wasfiltered through celite. The filtrates were concentrated and purified onsilica gel to give 104 g, 772RB147B as an oil.

To a solution of 772RB147B (67.4 g, 107 mmol), MPMCl (21.9 mL, 161 mmol,1.5 eq.) and a 1M solution of NaHMDS in THF (140 mL, 140 mmol, 1.3 eq.)was added slowly with syringe pump in 2 h at 0° C. After stirred for 1.5h at 0° C., it was quenched with sat. NH₄Cl at 0° C. and warmed to rt.The mixture was extracted with EtOAc (3×). The extracts were washed withwater, brine, dried and concentrated. The crude product was purified onsilica gel to give 772RB162 quantitatively.

772RB162 (119.6 g, 160 mmol) was dissolved in a mixture of 10% NaOH inmethanol (3.2 L, v/v) and 3.5 mL of water. The reaction was heated at45° C. for 48 h. After cooled, it was diluted with 9 L of CH₂Cl₂, washedwith water (2×), Sat NH₄Cl, brine, dried and concentrated. The crudeproduct was purified on silica gel with 10%-25-35% of EtOAc/Hexanes togive the 772RB164 (78 g, 96% yield).

To a solution of (COCl)₂ (25 mL, 295 mmol, 2 eq.) in CH₂Cl₂ (870 mL),DMSO (41.85 mL, 590 mmol, 4 eq.) was slowly added at −78° C. After 30min stirred at that temperature, a solution of 772RB164 (75 g, 147.4mmol) in CH₂Cl₂ (160 mL) was added in 45 min. After stirred at −78° C.for 45 min, Et₃N (82.2 mL, 590 mmol, 4 eq.) was added at thattemperature. After stirred for 30 min, it was warmed to 0° C. for 1.5 h.The reaction was quenched with 750 mL of saturated NH₄Cl and extractedwith EtOAc (3×). The extracts were dried and concentrated. The crudeproduct was re-suspended with 2.5 L of 1:1 solution of EtOAc/Hexanes,washed with water (3×), brine, dried and concentrated. The crude product772RB169 was used directly for next step.

To a suspension of Ph₃PCH₃Br (115.8 mL, 324.3 mmol, 2.2 eq.) in amixture of THF (870 mL) and DMSO (433.6 mL), n-BuLi (184.3 mL of 1.6 Msolution, 294.8 mmol, 2 eq.) was added at 0° C. After stirred for 1 h, asolution of 772RB169 (74.7 g in 175 mL of THF, 147.4 mmol) was added at0° C. After 30 min, it was warmed to rt. After 2 h, it was quenched with1.1 L of Sat. NHCl₄ and extracted with EtOAc (3×). The extracts werewashed with water, brine and dried and concentrated. The crude productwas purified on silica gel with 5-10% EtOAc/Hexanes to give 66.5 g of772RB170 as an oil (89% yield).

3) Coupling of Triflate and Olefin:

To a mixture of 772RB168 (2.5 g, 4.95 mmol) and triflate (2.7 g, 6.4mmol, 1.3 eq.), Pd₂(dba)₃ (1.36 g, 1.48 mmol, 0.3 eq.) was added in thedry box. After moved out of dry box, the mixture was suspended in 8.3 mLof dioxane and N-Methyl N-dicyclohexane amine (2.1 mL, 9.9 mmol, 2 eq.)was added. The reaction was heated at 100° C. for 12 h with vigorousstirring. After cooled, 6 g of celite was added and diluted with EtOAc.The mixture was filtered through a celite plug and rinsed with EtOAc.The filtrates were concentrated. The crude product was purified onsilica gel with 10-20% EtOAc/Hexanes to give 3 g of pure 772RB172 (76%yield).

To a solution of 772RB173 (46.3 g, 58.16 mmol) in DMPU (291 mL), LiHMDS(116 mL of 1M solution in THF, 116.3 mmol, 2 eq.) was added at 0° C.After stirred for 40 min at that temperature, EtI (27.9 mL, 349 mmol, 6eq.) was added. After 5 min, it was warmed to rt. After stirred 3 h, itwas quenched with 1 L of Sat. NHCl4 at 0° C. The mixture was extractedwith MTBE/Hexanes (1:1). The extracts were washed with brine, dried andconcentrated. The crude product was purified on silica gel with 15-20%EtOAc/Hexanes to give 40 g of desired product, 77RB175 (84% yield).

To a solution of 772RB175 (48 g, 58.2 mmol) in 230 mL, a solution ofTBAF (407 mL of 1M solution, 407 mmol, 7 eq.) and imidazole.HCl (21.3 g,203.9 mmol, 3.5 eq.) was added. The reaction was heated at 60° C. for 72h. After cooled to rt, it was quenched with sat. NH₄Cl and was extractedwith ether (3×). The organic layers were washed with water, brine, driedand concentrated. The crude product was purified on silica gel with20-30% EtOAc/Hexanes to give 31.4 g (76% yield).

To a solution of 772RB177 (20.3 g, 28.6 g) in 3 L of THF, 0.5M KHMDSsolution (60 mL, 30 mmol, 1.05 eq.) was added slowly by syringe pump in120 min. After stirred for 5 min, it was quenched with 1.5 L of sat.NH₄Cl. The mixture was extracted with ether (3×). The extracts werewashed with brine, dried and concentrated. The crude product waspurified on silica gel with 10-20%-50% EtOAc/Hexanes to give 14.2 g ofdesired product (76% yield).

To a solution of 772RB178, 179 (19 g, 29.15 mmol) in DMF (194 mL),imidazole (4 g, 58.3 mmol, 2 eq.) and TBSCl (5.27 g, 35 mmol, 1.2 eq.)were added. After stirred overnight, it was quenched with a saturatedsolution of NaHCO₃ and water. The mixture was extracted with EtOAc. Theorganic layer was washed with, water, brine, dried and concentrated. Thecrude product was purified on silica gel column to give 22 g (99% yield)of desired product.

To a solution of 772RB181 (22 g, 28.7 mmol) in a mixture of CH₂Cl₂ (230mL) and H₂O (57.4 mL), DDQ (9.78 g, 43 mmol, 1.5 eq.) was added at 0° C.After stirred for 2 h, it was quenched with 1 L of a 1:1 mixture of aq.saturated NaHCO₃ and 10% aq. Na₂S₂O₃. The mixture was extracted with 3×1L of ether. The extracts were washed with brine, dried and concentrated.The crude product was purified on silica gel to give 18.1 g of pureproduct.

To a solution of 772RB182 (18 g, 27.9 mmol) in 279 mL of CH₂Cl₂, dried 4Å molecular sieves (18 g) and PCC (18 g) were added. After stirred for90 min, it was quenched with Et₃N (19.45 mL). The reaction mixture wasfiltered through a plug of celite, the plug was rinsed with 75% EtOAc inhexanes (900 mL). The filtrates were concentrated. The crude product waspurified on silica gel column with 10-15-20% EtOAc/Hexanes to give 14.6g (81%) pure product.

In a 2 L flask, 772RB183 (8.5 g, 13.2 mmol) was dissolved in CH₂Cl₂(82.5 mL) and the mixture was cooled to 0° C. Then a solution of 5%H₂O/TFA (4.1/78.1 mL) was added slowly (˜30 min) and the mixture wasstirred at 0° C. for 14.5 hrs. The reaction was monitored by TLC.Reaction mixture was diluted with CH₂Cl₂ at 0° C. Reaction was quenchedwith a solution of NaHCO₃ in water (˜1.2 eq compare to TFA). Reactionwas cooled to r.t. Extract 3× with CH₂Cl₂, dried with Na₂SO₄, Filtered,and concentrated. Chromatography on Si-Gel, 50-60-75% EtOAc/hexane gaveER-806201: 4.8 g, 93% yield.

Preparation of C5-F-Enone Series:

Preparation of ER803030:

496-SG-026B

To a magnetically stirred solution of 2-fluoro-2-phosphonoacetic acidtriethylester (8 g, 33.3 mmol) in DMF (2.7 mL) at 0° C. was introducedsodium hydride (0.8 g, 33.3 mmol). After 1 hour of stirring at 0° C., asolution of aldehyde (3.47 g, 16.65 mmol) in THF (14 mL) was added.After 2.5 hours of stirring at 0° C., a saturated solution of ammoniumchloride was added. The reaction mixture was diluted with water andextracted with ethyl acetate. The crude product was purified by flashchromatography eluting with n-hexane/ethyl acetate: (20/1) to afford496-SG-026B (3.58 g, 72% yield).

496-SG-027B

To a magnetically stirred solution of 496-SG-026B (3.58 g, 12.1 mmol) indichloromethane (136 mL) at 0° C. was introduced DIBAL-H (1M solution indichloromethane, 30.2 mL, 30.2 mmol). After 0.5 hour of stirring at 0°C., the reaction mixture was warmed-up to room temperature and stirredan additional 10 minutes. The reaction was cooled back to 0° C. and asaturated solution of ammonium chloride was added (5.4 mL). After 15minutes of stirring, the reaction mixture was diluted with ether andstirred 30 minutes at room temperature. The resulting suspension wasfiltered and the solid washed with ether. The solvent was removed byevaporation. The crude product was purified by flash chromatographyeluting with hexanes/ethyl acetate: (3/1) to afford 496-SG-027B (2.68 g,87% yield).

496-SG-028B

To a magnetically stirred solution of 496-SG-027B (2.68 g, 10.55 mmol)in dichloromethane (53.2 mL) cooled to 0° C. (Ice/water, externalthermometer) was introduced DMSO (2.6 mL, 36.94 mmol) followed by P₂O₅(5.24 g, 36.94 mmol). After one hours of stirring at room temperaturethe reaction was cooled down to 0° C., and triethylamine (7.4 mL, 52.72mmol) was added. After 20 minutes of stirring at room temperature thereaction mixture was diluted with water and extracted withdichloromethane. The solvent was removed by evaporation. The crudeproduct was purified by flash chromatography eluting with hexanes/ethylacetate: (3/1) to afford 496-SG-028B (2.66 g).

496-SG-022B

To a magnetically stirred solution of 3-butyn-1-ol (3.0 g, 42.8 mmol)and imidazole (14.6 g, 214 mmol) in dichloromethane (113 mL) at roomtemperature was introduced tert-butyldiphenylsilyl chloride (11.7 mL).After 18 hours of stirring at room temperature the reaction was dilutedwith water and extracted with ether. The solvent was removed byevaporation. The crude product was purified by flash chromatographyeluting with hexanes/ethyl acetate: (2/1) to afford 496-SG-022B (13.7g).

496-SG-029B

To a magnetically stirred solution of 496-SG-022B (6.5 g, 21.8 mmol) inTHF (208 mL) at −78° C. was introduced n-BuLi (2.5 M in hexane, 8.4 mL,21.1 mmol). After 1 hours of stirring −78° C., a solution of 496-SG-028B(2.66 g, 10.55 mmol) in THF (128 mL) was added at −78° C. After 15minutes of stirring at −78° C., the reaction was quenched by addition ofa saturated solution of ammonium chloride. The reaction mixture wasdiluted with water and extracted with ethyl acetate. The solvent wasremoved by evaporation. The crude product was purified by flashchromatography eluting with hexanes/ethyl acetate: (5/1) to afford496-SG-029B (4.27 g, 70%).

496-SG-30A

A solution of 496-SG-29B (4.27 g, 7.61 mmol) and quinoline (0.033 mL) inhexanes with a catalytic amount of Lindlar catalyst was magneticallystirred for 1 h under hydrogen atmosphere. The reaction mixture wasfiltered through celite, and the solvent removed by evaporation toafford 496-SG-30A (4.28 g). The crude was used in the next step withoutpurification.

496-SG-031B

To a magnetically stirred solution of 496-SG-030A (4.28 g 7.61 mmol),triethylamine (2.7 mL, 19.3 mmol) and a catalytic amount of DMAP indichloromethane (267 mL) at room temperature, was introduced benzoylchloride (1.8 mL, 15.22 mmol). After 18 hours of stirring at roomtemperature the reaction mixture was diluted with a 0.1 M solution ofsodium hydroxide and extracted with ether. The solvent was removed byevaporation. The crude product was purified by flash chromatographyeluting with hexanes/ethyl acetate: (9/1) to afford 496-SG-031B (5.0 g,98%).

496-SG-042B

To a magnetically stirred solution of 496-SG-031B (3.79 g, 5.68 mmol) inacetone (57 mL) at room temperature, was introduced NMO (1.33 g, 11.36mmol) and water (2.9 mL). The reaction mixture was cooled down to 0° C.,and a solution (0.1 M in toluene) osmium tetraoxide was added. After 18hours of stirring at room temperature the reaction was quenched withsodium thiosulfate and stirred at room temperature for 20 minutes. Thereaction mixture was then diluted with water and extracted with ethylacetate. The solvent was removed by evaporation. The crude product waspurified by flash chromatography eluting with hexanes/ethyl acetate:(3/1) to afford 496-SG-042B (1.5 g, 39%).

496-SG-043B

To a magnetically stirred solution of 496-SG-042B (2.16 g, 3.08 mmol)and 2,2-dimethoxypropane (1.93 mL, 15.4 mmol) in a 2/1 mixtureacetone/dichloromethane (32 mL) at room temperature, was introducedcamphosufonic acid (0.8 g, 3.4 mmol). After 2 hours of stirring at roomtemperature the reaction was quenched with sodium bicarbonate and thereaction mixture was then diluted with water and extracted with ethylacetate. The solvent was removed by evaporation. The crude product waspurified by flash chromatography eluting with hexanes/ethyl acetate:(5/1) to afford 496-SG-043B (2.13 g, 93%).

496SG-45A

To a magnetically stirred solution of 496-SG-043B (2.14 g, 2.89 mmol) inTHF (52 mL) at 0° C., was introduced a 1 M solution of TBAF in THFbuffered with 0.5 equivalent of imidazole hydrochloride (7.2 mL, 7.2mmol). After 1 hour of stirring at room temperature the reaction wasdiluted with water and extracted with ether. The solvent was removed byevaporation to afford 496SG-45A (1.39 g). The crude product was used inthe next step without purification.

496-SG-046B

Using a procedure analogous to that described for the synthesis of343-YW-281, 496-SG-045A (1.4 g, 2.79 mmol) was reacted withtriphenylphosphine (1.24 g, 4.74 mmol), DEAD (0.465 mL, 2.93 mmol) andmethyl iodide (0.225 mL, 3.63 mmol) in toluene (46.5 mL). The crudeproduct was purified by flash chromatography eluting with hexanes/ethylacetate (5/1 and then 3/1) to afford 496-SG-046B (1.46 g, 85% yield).

496-SG-048B

Using a procedure analogous to that described for the synthesis ofER-803027 (stage 447-JCH-273B), 496-SG-046B (1.46 g, 2.38 mmol) wasreacted with intermediate 509-HD-213 (178 g, 3.00 mmol) and LiHMDS (1Msolution in THF, 3.6 mL, 3.6 mmol) in a 10/1 THF/HMPA mixture (17.3 mL)to afford after purification by flash chromatography eluting withhexanes/ethyl acetate: 496-SG-048A. Using a procedure analogous to thatdescribed for the synthesis of 447-JCH-275B, 496-SG-048A was reactedwith MCPBA (0.75 g, 2.38 mmol) and triethylamine (2 mL, 14.3 mmol). Thecrude product was purified by flash chromatography eluting withhexanes/ethyl acetate (5/1 and then 3/1) to afford 496-SG-048B (1.38 g,72% yield).

496-SG-052B

496-SG-48B (1.28 g, 1.58 mmol) was reacted with DDQ (0.43 g, 1.89 mmol)in a 2/1-dichloromethane/water mixture (68 mL). The crude product waspurified by flash chromatography eluting with hexanes/ethyl acetate (5/1and then 3/1) to afford 496-SG-052B (0.88 g, 81% yield).

496-SG-053A

Using a procedure analogous to that described for the synthesis ofER-803064 (stage 509-HD-116), 496-SG-052B (0.88 g, 1.28 mmol) wasreacted with TBAF (1.0 g, 3.82 mmol) in THF (2.4 mL) to afford496-SG-053A (0.74 g). The crude product was used in the next stepwithout purification.

496-SG-058B

496-SG-053A (0.20 g, 0.34 mmol) was reacted with triphenylphosphine(0.107 g, 0.408 mmol) and DEAD (0.064 mL, 0.408 mmol) in THF (90 mL).The crude product was purified on silica gel eluting with n-hexane/ethylacetate: (2/1) to afford 496-SG-058B (0.12 g, 62% yield).

496-SG-057A, 496-SG-057B, 496-SG-057C

Using a procedure analogous to that described for the synthesis ofER-803064, 496-SG-058B (0.048 g, 0.084 mmol) was reacted with sodiumhydroxide (1M solution, 0.42 mL, 0.42 mmol) in a 2/1 mixture ethanol/THF(1 mL). The crude product was purified on silica gel (TLC) eluting withhexanes/ethyl acetate: (1/1) to afford 496-SG-057A (0.011 g),496-SG-057B (0.013 g), 496-SG-057C (0.01 g).

496-SG-061A, 496-SG-061B, 496-SG-061C.

496-SG-057A (0.01 g, 0.021 mmol), 496-SG-057B (0.012 g, 0.026 mmol),496-SG-057C (0.0095 g, 0.02 mmol) were separately reacted withDess-Martin reagent: (0.055 g, 0.129 mmol), (0.065 g, 0.154 mmol),(0.052 g, 0.122 mmol) and sodium carbonate (0.027 g), (0.032 g), (0.026g) in dichloromethane (1.5 mL), (1.8 mL), (1.4 mL). After work-up, eachreaction mixture was purified on silica gel (TLC) eluting withhexanes/ethyl acetate: (2/1) to afford respectively 496-SG-061A (0.009g), 496-SG-061B (0.006 g), and 496-SG-061C (0.011 g).

496-SG-067A/ER-803029, 496-SG-067B/ER-803026, 496-SG-067C/ER-803030.

Using a procedure analogous to that described for the synthesis ofER-803064 (final step), 496-SG-061A (0.007 g, 0.016 mmol), 496-SG-061B(0.004 g, 0.009 mmol), and 496-SG-061C (0.013 g, 0.027 mmol) wereseparately reacted with HF 48%: (0.37 mL), (0.21 mL), (0.63 mL) inacetonitrile/dichloromethane (4/1):(1.9 mL), (1.0 mL), (3 mL). Afterwork-up, each reaction mixture was purified on silica gel (TLC) elutingwith hexanes/ethyl acetate: (2/1) to afford respectively496-SG-067A/ER-803029 (0.006 g), 496-SG-067B/ER-803026 (0.002 g), and496-SG-067C/ER-803030 (0.006 g).

ER803916

To a solution of (EtO)₂POCHFCO₂Et (5.6 mL) in a mixture of THF and DMF(100 and 25 mL), NaH (1.2 g) was added at 0° C. After stirred at 0° C.for 1 h, it was cooled to −40° C. with dry ice-acetone bath. A solutionof aldehyde (3.5 g) in THF (20 mL) was added drop wise. Then, thereaction was warmed up to rt overnight. It was quenched with sat. NH₄Cl,and extracted with EtOAc (2×). The organic layers were washed withbrine, dried and concentrated under vacuum. The crude product waspurified on silica gel with 15:1, Hexanes:EtOAc to give 3.5 g of desiredproduct, 531-yw-11, which gave satisfactory ¹H NMR. The cis:trans ratioof the two isomers was 12:1 based on ¹H NMR.

To a solution of ester, 531-W-11, (3.5 g) in ether (300 mL), a solutionof DIBAL-H (15 mL) was added at 0° C. After 1 h at 0° C., it wasquenched with 4 mL of MeOH, and 15 mL of aq. Sat. Na₂SO₄. After stirredfor 51 hr at rt, it was filtered through a pad of celite, and the padwas washed with ether (2×). The combined filtrates were concentrated todryness to give 3.5 g of crude product, 531-YW-12.

To a solution of alcohol (5.6 g) and MPMOTCl (24 g) in 120 mL of Et₂O, asolution of TfOH (20 mL, 0.3 mL dissolved in 25 mL of Et₂O) was added in4 h. Then it was quenched with Sat. NaHCO₃ and extracted with Et₂O (2×).The organic layers were washed with brine, dried and concentrated. Thecrude solid was suspended with pentanes. The precipitation was filtered.The filtrates were concentrated to give 15 g of crude product. It waspurified on silica gel with 8:1, Hexanes/EtOAc to give 12.7 g of thedesired product, 531-YW-14

To a solution of (COCl)₂ (5 mL) in 250 mL of CH₂Cl₂, DMSO (10 mL) wasadded at −78° C. After 15 min at −78° C. a solution of 531-YW-12 (3.8 g)in 50 mL of CH₂Cl₂ was added at that temperature. After 30 min at −78°C., Et₃N (15 mL) was added, the reaction was warmed to 0° C. Thereaction was quenched with sat. NH₄Cl, extracted with EtOAc. The organiclayer was washed with brine, dried and concentrated. The crude productwas passed through a short silica gel pad with Hexanes/EtOAc, 8:1 togive 4.1 g of slightly impure product.

To a solution of acetylene, 531-YW-14 (3.56 g, 18.7 mmol, 1 eq.) in 200mL of THF, a solution of n-BuLi (12.9 mL, 20.58, 1.1 eq., 1.6 mmol) wasadded at −78° C. The reaction was warmed to 0° C. for 5 min (by internaltemperature). Then it was cooled back to −78° C. A solution of aldehyde,531-YW-12 (9.8 mmol, 0.5 eq.) in 50 mL of THF was added. The reactionwas allowed to be warmed up to 0° C. in 1 h. It was quenched with sat.NH₄Cl and purified as described before to give the desired acetylenicalcohol, 531-YW-15.

The starting material (531-YW-15, 5.7 g, 10.2 mmol) was dissolved in 200mL hexanes. Quinoline (500 μL) and Lindlar catalyst (1.0 g) were added.The reaction mixture was stirred at room temperature under H₂ balloonatmosphere for 1 h. Then the catalyst was filtered away. Quantitativeamount of 509-HD-134 was obtained as colorless oil.

509-HD-134 (5.7 g, 10.2 mmol) was dissolved in 100 mL dichloromethane atroom temperature. Triethylamine (3.5 mL, 25.5 mmol), benzoyl chloride(2.4 mL, 20.4 mmol) and catalytic amount of DAMP were added,respectively. After stirring for 1 h, 0.1N sodium hydroxide solution wasadded and the reaction mixture was extracted with ethyl acetate. Thecrude product was purified on silica gel column, giving 509-HD-135 ascolorless oil in 77% yield.

509-HD-135 (5.2 g, 7.64 mmol) was dissolved in acetone and water(1:0.05) at 0° C. 4-Methylmorpholine N-oxide (1.8 g, 15.28 mmol) andsolution of osmium tetraoxide (0.1M, 7.6 mL) in toluene were added. Thereaction mixture was warmed up to room temperature and stirred for 20 h.It was quenched with 10% sodium thiosulfate in sat. sodium bicarbonateaqueous solution, and extracted with ethyl acetate. After purificationon silica gel column, 509-HD-138 was obtained in 93% yield.

509-HD-138 (5.1 g, 7.13 mmol) was dissolved in 80 mL of dichloromethane.2-Methoxypropene (1.4 mL, 14.26 mmol) and catalytic amount of pyridiniump-toluenesulfonate were added. After stirring at room temperature for 20min, the reaction mixture was quenched with sat. sodium bicarbonatesolution and extracted with dichloromethane. After purification onsilica gel column, 509-HD-139 was obtained in 90% yield.

509-HD-139 (2.55 g, 3.38 mmol) was dissolved in the mixture of 30 mL ofdichloromethane and 15 mL of water.2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (844 mg, 3.72 mmol) was added.After stirred at room temperature for 1 h, the reaction mixture wasquenched with sat. sodium bicarbonate solution and extracted with ethylacetate. After purification on silica gel column, 509-HD-140 wasobtained as white foam in 98% yield.

509-HD-140 (1.86 g, 2.93 mmol) was dissolved in 30 mL of toluene at roomtemperature. Triphenylphosphine (1.31 g, 4.98 mmol) was added, followedby methyl iodide (236 μL, 3.80 mmol) and diethyl azodicarboxylate (508L, 3.22 mmol). After stirring for 10 min, the reaction mixture wastriturated with toluene. After purification on silica gel column,509-HD-141 was obtained as colorless oil in 96% yield.

509-HD-141 (2.03 g, 2.73 mmol) and 509-HD-213 (2.84 g, 5.46 mmol) weredissolved in a mixture of 35 mL of THF/HMPA (10/1) at −78° C. Thesolution of LiHMDS (1N, 4.6 mL) in hexanes was added. The reactionmixture was stirred for 30 min, and then it was quenched with sat.ammonium chloride and extracted with ethyl acetate. After purificationon silica gel column, 509-HD-143 was obtained in 85% yield.

509-HD-143 (2.85 g, 2.59 mmol) was dissolved in 50 mL of dichloromethaneat 0° C. 3-Chloroperbenzoic acid (50%, 1.8 g) was added. After stirringat 0° C. for 20 min, triethylamine (2.2 mL, 15.5 mmol) was added, thenthe reaction mixture was warmed up to room temperature and stirred for30 min. It was quenched with 10% sodium thiosulfate in sat. sodiumbicarbonate aqueous solution, and extracted with dichloromethane. Afterpurification on silica gel column, 509-HD-144 was obtained as white foamin 63% yield.

509-HD-144 (1.33 g, 1.41 mmol) was dissolved in 10 mL of THF. The THFsolution of TBAF buffered with imidazole.HCl (1N, 14.1 mL) was added.The reaction mixture was heated at 50° C. for 72 h. It was diluted withEt₂O and washed with H₂O. After purification on silica gel column,509-HD-150 was obtained as pale yellow foam in 95% yield.

2-Chloro-1-methylpyridinium iodide (1.03 g, 4.02 mmol) and n-Bu₃N (958μl, 4.02 mmol) were dissolved in 80 mL of dichloromethane and heated toreflux. The solution of 509-HD-150 (807 mg, 1.34 mmol) in 50 mL ofdichloromethane was added slowly. The reaction mixture was heated for 30min. It was washed with 0.02N hydrochloric acid, sat. sodium bicarbonatesolution and brine, respectively. After purification on silica gelcolumn, 509-HD-152 was obtained in 50% yield.

509-HD-152 (390 mg, 0.67 mmol) was dissolved in 10 ml ethyl alcohol.Sodium hydroxide (1N, 6.7 mL) solution was added. The reaction mixturewas stirred for 1 h at room temperature. It was diluted with H₂O,extracted with EtOAc. After purification on silica gel column, 134 mg ofthe major desired single isomer 509-HD-153C was obtained as colorlessoil.

509-HD-153C (94.4 mg, 0.20 mmol) was dissolved in 8 mL ofdichloromethane. Molecular sieve (4 A, 423 mg) and PCC (423 mg, 2.0mmol) were added. The reaction mixture was stirred for 12 h at roomtemperature. After passing through celite, 509-HD-158 was obtained ascolorless oil in 52% yield.

509-HD-158 (49.2 mg, 0.10 mmol) was dissolved in 1.0 mL ofdichloromethane. Then hydrofluoric acid (6N, 4 mL) was added. Thereaction mixture was stirred at room temperature for 30 min. It wasdiluted with more dichloromethane, washed with water and sat. sodiumbicarbonate solution. After purification on a plug of silica gel,ER803916 was obtained as a white solid in 81% yield.

Preparation of Intermediate for the Synthesis of ER806821:

The C14-TMS-ethyl intermediate was prepared from C4-TMS ethyl selenidein analogous to the above sequences in similar yields.

The phenol was prepared using previous describe conditions for the C14modification series:

Preparation of ER806821:

To a solution of phenol (18.8 g, 33 mmol) in CH₂Cl₂ (300 mL) at 0° C.was added Et₃N (11.5 mL, 82.5 mmol). Then, Tf₂O (8.3 mL, 49.5 mmol) inCH₂Cl₂ (35 mL) was added dropwise over a period of 60 min and thereaction was stirred for an additional 30 min. The reaction was quenchedat 0° C. with a saturated solution of NaHCO₃ (200 mL), extracted 3 timeswith 300 mL of CH₂Cl₂, the combined organic layers were dried withNa₂SO₄, the solid was filtered and the solvent was evaporated undervacuo. The crude compound was purified by flash chromatography on silicagel using 10% EtOAc/hexane as eluent to give 20.1 g (28.6 mmol, 87%) of640-RB-297 as white foam.

In a glove box, under N₂, a 200 mL round-bottom flask equipped with amagnetic stirring bar was charged with 640-RB-297 (3.0 g, 4.27 mmol),N-Methyl-N-Boc amine (784 mg, 5.98 mmol),Tris(dibenzylideneacetone)-dipalladium (196 mg, 0.21 mmol),2-di-t-butylphosphino-biphenyl (127 mg, 0.427 mmol), sodium-t-butoxide(574 mg, 5.98 mmol) and anhydrous toluene (100 mL). The flask wasremoved from the glove box and the mixture was stirred at 80° C. for 4hrs. Then the mixture was allowed to cool to room temperature, 100 mL ofa saturated solution of NH₄Cl was added and the mixture was stirred for1 hr. The mixture was extracted 3 times with 100 mL of EtOAc, theorganic layers were mixed together, dried with Na₂SO₄, the solid wasfiltered through a short plug of Si-Gel, rinsed with EtOAc and thesolvent was evaporated under vacuo. The crude compound was purified byflash chromatography on silica gel using 20% EtOAc/hexane as eluent togive 1.67 g (2.44 mmol, 57%) of 772-RB-20 as white foam.

To a solution of 772-RB-20 (1.67 g, 2.44 mmol) in EtOH (75 mL) and THF(5 mL) was added a solution of 1N NaOH (24.4 mL, 24.4 mmol) and theresulting mixture was stirred at room temperature for 4 hrs. Then, EtOHwas partially evaporated, the mixture was diluted with 500 ml EtOAc,washed with water (2×250 mL) and brine (250 mL), the organic phase wasdried with Na₂SO₄, the solid was filtered and the solvent wasevaporated. The crude compound was purified by flash chromatography onsilica gel using 20-30% EtOAc/hexane as eluent to give 620 mg (1.07mmol, 44%) of 772-RB-21A and 370 mg (0.64 mmol, 26%) of 772-RB-21B(undesired stereochemistry at C8-C9) as white foams.

To a solution of 772-RB-21A (610 mg, 10.05 mmol) in CH₂Cl₂ (25 mL) wasadded pyridine (0.425 mL, 5.25 mmol) and then Dess-Martin periodinane(1.10 g, 2.6 mmol). The mixture was stirred at room temperature for 1.5hr. It was then diluted with 100 mL of Et₂O, the solid was removed byfiltration through celite and rinsed with 100 mL of Et₂O. The organicextract was dropped in a solution of 10% w/v Na₂S₂O₄ in saturated NaHCO₃solution and stirred for 15 min. The phases were separated, the organiclayer was washed with 100 mL of brine, dried with Na₂SO₄, the solid wasfiltered and the solvent was evaporated. The crude compound was purifiedby flash chromatography on silica gel using 30% EtOAc/hexane as eluentto give 560 mg (0.97 mmol, 92%) of 772-RB-22 as white foam. The otherdiastereomer 772-RB-21B was treated the same way to give 325 mg (0.56mmol, 91%) of 772-RB-25 as white foam.

A solution of 772-RB-22 (140 mg, 0.242 mmol) in CH₂Cl₂ was cooled downto −35° C. in a bath of dry ice/acetone. Then a solution of 5% H₂O/TFAwas added slowly. The mixture was warmed up slowly to −23° C. andstirred at this temperature for 50 min. Then the mixture was cooled downto −35° C., neutralized with a saturated solution of NaHCO₃ and allowedto warm up to room temperature The mixture was extracted 3 times withCH₂Cl₂, the combined organic layers were dried with Na₂SO₄, the solidwas filtered and the solvent evaporated. This reaction was repeated 4times. The crude compound was purified by flash chromatography on silicagel using 40% EtOAc/hexane as eluent to give 161 mg (0.42 mmol, 42%) ofER-806821. The other diastereomer 772-RB-25 was deprotected the same wayand then the C₈-C₉ diol was isomerized by treatment with H₂O and Silicagel in CH₂Cl₂ overnight to give 70 mg (0.178 mmol, 32% 2 steps) ofER-806821.

Alternative Synthesis of ER806821 and Synthesis of ER807563:

Synthesis of Acyclic Intermediate 20:

Preparation of Aldehyde 29:

Aldehyde 29 was prepared according to the procedure describedpreviously.

Preparation of Alkyne 30:

To a solution of 3-butyn-1-ol (8.98 g, 128 mmol, 1.0 eq.) indichloromethane (200 mL) was added triethylamine (23.5 mL, 167 mol, 1.3eq.) and 4-(dimethylamino)-pyridine (25 mg, 0.205 mmol, 0.0016 eq.). Theresulting mixture was cooled down to 0° C., added slowlytrimethyl-acetyl chloride (17.5 mL, 141 mmol, 1.1 eq.). Once exothermwas over, the reaction mixture was allowed to warm up to rt. Stirredovernight at rt. A saturated solution of NaHCO₃ was added, the biphasicmixture was stirred at rt during 1 h. Layers were separated. The aqueousone was extracted with dichloromethane. The organic layers were combinedand dried with sodium sulfate, filtered and concentrated to dryness. Thecrude oil was purified by filtration on a pad of silica gel 230-400 Meshusing 20% ethyl acetate/hexane to elute. This procedure afforded alkyne30, 17.3 g, pale yellow oil, 88% yield.

Preparation of Carbinol 31:

Zinc triflate (1.73 g, 7.31 mmol, 1.1 eq.) and(1S,2R)-(+)-N-methylephedrine (1.43 g, 7.90 mmol, 1.2 eq.) were added toa 100 mL round bottom flask in a dry box. The flask was transferred to afume hood for the addition of toluene (20 mL) andN,N-diisopropylethylamine (1.39 ml, 7.94 mmol, 1.2 eq.). The resultingmixture was stirred 2 h at rt at which point alkyne 30 (1.23 g, 7.97mmol, 1.2 eq.) was added. Stirred 60 min at rt, added aldehyde 29 (1.73g, 6.64 mmol, 1.0 eq.) and stirred at rt 40 min. Quenched by theaddition of a saturated solution of ammonium chloride. Layers wereseparated, the aqueous one was extracted with Et₂O, the organic layerswere combined, washed with brine, dried with sodium sulfate, filteredand concentrated to dryness. The resulting crude oil was purified bychromatography on silica gel 230-400 Mesh using the following solventgradient: 8%, 12% and 16% ethyl acetate/hexane to elute. This procedureafforded carbinol 31, 2.21 g, colorless oil, 80% yield, 94% de.

Preparation of Alkene 32:

To a solution of carbinol 31 (2.60 g, 6.27 mmol, 1.0 eq.) in n-heptane(70 mL) was added quinoline (0.16 mL, 1.33 mmol, 0.21 eq.) and Lindlarcatalyst (640 mg). The resulting heterogeneous mixture was stirred at rtduring 1.5 h under a positive atmosphere of hydrogen. After that time,the reaction was found complete. It was then filtered on Celite and thefiltrate was washed with a dilute HCl solution (prepared by mixing HCl 1N (6.0 mL) with water (24 mL)) to remove quinoline. Layers wereseparated, the organic one was washed with water (3×20 mL), dried withsodium sulfate, filtered and concentrated to dryness to afford alkene32, 2.56 g, colorless oil, 98% yield.

Preparation of Ether 33:

To a solution of alkene 32 (2.40 g, 5.76 mmol, 1.0 eq.) and thetrichloroimidate of 4-methoxybenzyl alcohol (2.05 g, 7.20 mmol., 1.25eq.) in dichloromethane (7.5 mL) was added at rt pyridiniump-toluenesulfonate (74 mg, 0.29 mmol, 0.05 eq.). The reaction mixturewas stirred at rt overnight. The reaction was found incomplete and inorder to push it to completion it required sequential additions of extrapyridinium p-toluenesulfonate (74 mg, 0.29 mmol, 0.05 eq.) andtrichloroimidate of 4-methoxy-benzyl alcohol (2.05 g, 7.20 mmol., 1.25eq.) and prolonged stirring at 40° C. The reaction was quenched byadding a 1:1 mixture of THF and water (10 mL) and stirring was continuedfor 1 h at 40° C. Dichloromethane was added, layers were separated, theorganic one was washed with NaHCO₃, dried with sodium sulfate, filteredand concentrated to dryness. The crude oil was purified bychromatography on silica gel 230-400 Mesh using the following solventgradient: 5% and 7.5% ethyl acetate/hexane to elute. This procedureafforded ether 33, 2.60 g, colorless oil, 84% yield.

Preparation of Diol 34:

To a solution of ether 33 (2.20 g, 4.10 mmol, 1.0 eq.) in THF (28 mL)was added a solution of 4-methyl morpholine N-oxide (990 mg, 8.20 mmol,2.0 eq.) in water (28.0 mL). The mixture was added at rt a solution ofosmium tetroxide (0.55 mL, 0.055 mmol, 0.013 eq., osmium tetroxide 0.1 Min water). The reaction mixture was stirred at rt during 19.5 h. Then itwas quenched by the addition of 20 mL of a 1:1 solution of NaHCO₃saturated and 10% Na₂S₂O₃ in water. Stirring was continued during 60 minat rt. Layers were separated; the aqueous one was extracted with ethylacetate. The organic layers were combined, washed with brine, dried withsodium sulfate, filtered and concentrated to dryness. The resultingcrude oil was purified by chromatography on silica gel 230-400 Meshusing the following solvent gradient: 15% and 40% ethyl acetate/hexaneto elute. This procedure afforded diol 34, 2.06 g, colorless oil, 88%yield beta/alpha ratio>=10:1.

Preparation of Acetonide 35:

To a solution of diol 34 (2.17 g, 3.80 mmol, 1.0 eq.) and2-methoxypropene (0.77 mL, 7.64 mmol, 2.0 eq.) in dichloromethane (40mL) was added at rt pyridinium p-toluenesulfonate (10 mg, 0.039 mmol,0.01 eq.). The reaction mixture was stirred at rt during 45 min andquenched by the addition of a saturated solution NaHCO₃. Dichloromethanewas added, layers were separated, the aqueous one was extracted withdichloromethane, the organic layers were combined, washed with brine,dried with sodium sulfate, filtered and concentrated to dryness. Thecrude oil was purified by chromatography on silica gel 230-400 Meshusing 10% ethyl acetate/hexane to elute. This procedure affordedacetonide 35, 1.80 g, a colorless oil, 78% yield.

Preparation of Alcohol 36:

To a solution of acetonide 35 (940 mg, 1.54 mmol, 1.0 eq.) in methanol(15 mL) at rt was added potassium carbonate (261 mg, 1.85 mmol, 1.2eq.). The resulting mixture was stirred at 50° C. during 21 h, cooleddown to rt, added n-hexane (30 mL), water (15 mL) and a saturatedsolution of NH₄Cl (30 mL). Layers were separated, the aqueous one wasextracted with ethyl ether/hexane, the organic layers were combined,dried with sodium sulfate, filtered and concentrated to dryness. Theresulting crude oil was purified by chromatography on silica gel 230-400Mesh using the following solvent gradient: 20%, 60% and 80% ethylacetate/hexane to elute. This procedure afforded alcohol 36, 787 mg, acolorless oil, 97% yield.

Preparation of Aldehyde 37:

To a solution of dimethyl sulfoxide (0.44 mL, 6.19 mmol, 4.0 eq.) indichloromethane (5.0 mL) at −78° C. was added oxalyl chloride (1.54 mL,3.08 mmol, 2.1 eq., oxalyl chloride 2.0 M in dichloromethane). Theresulting mixture was stirred 30 min at −78° C., added alcohol 36 (787mg, 1.49 mmol, 1.0 eq.) in dichloromethane (2.0 mL to dissolve alcoholand 2×1.5 mL to rinse flask). The reaction mixture was stirred 60 min at−78° C., added triethylamine (0.87 mL, 6.22 mmol, 4.2 eq.), stirred 5min at −78° C., then warmed up to rt and stirring was continued at thattemperature during 50 min. At that point a saturated solution of NH₄Clwas added. Layers were separated and the aqueous one was extracted threetimes with dichloromethane. The organic layers were combined, dried withsodium sulfate, filtered and concentrated to dryness. The crude oil wasdissolved in a 1:1 mixture of ethyl acetate/hexane (100 mL) and washedwith water (10 mL) three times, then washed with brine, dried withsodium sulfate, filtered and concentrated to dryness. The concentratedmaterial was azeotroped with toluene and pumped to constant weight toafford aldehyde 37, 791 mg, a colorless oil, quantitative yield.

Preparation of Alkene 20:

To a suspension of methyltriphenylphosphonium bromide (549 mg, 1.51mmol, 2.2 eq.) in dimethyl sulfoxide (1.0 mL) and THF (3.6 mL) at 0° C.was added n-butyllithium (0.855 mL, 1.37 mmol, 2.0 eq., n-butyllithium1.6 M in THF). The resulting mixture was stirred 60 min at 0° C., addedaldehyde 37 (359 mg, 0.684 mmol, 1.0 eq.) in THF (2.0 mL and 3×1.0 mLfor rinse). Stirred 20 min at 0° C., allowed to warm up to rt andstirred 1.5 h at that temperature. Quenched by the addition of asaturated solution of NH₄Cl, layers were separated and the aqueous onewas extracted with a 1:1 mixture of ethyl ether/hexanes. The organiclayers were combined, washed with brine, dried with sodium sulfate,filtered and concentrated to dryness. The resulting crude oil waspurified by chromatography on silica gel 230-400 Mesh using thefollowing solvent gradient: 5% and 6.5% ethyl acetate/hexane to elute.This procedure afforded alkene 20, 322 mg, a colorless oil, 90% yield.

B) Synthesis of ER-806821:

Preparation of Heck-Coupling Product 22:

Alkene 20 (380 mg, 0.727 mmol, 1.0 eq.) and triflate 21 (321 mg, 0.727mmol, 1.0 eq.) were combined and addedtris-(dibenzylideneacetone)-dipalladium-(0) (33.2 mg, 0.036 mmol, 0.05eq.) under inert atmosphere (dry box). Those reagents were moved out ofthe dry box and added at rt acetonitrile (3.0 mL) and triethylamine(0.205 mL, 1.46 mmol, 2.0 eq.). The resulting mixture was stirred at 65°C. during 19 h, cooled down to rt and filtered on Celite. The filtratewas concentrated to dryness and purified by chromatography on silica gel230-400 Mesh using the following solvent gradient: 5%, 15%, 20% and 30%ethyl acetate/hexane to elute. This procedure afforded Heck-couplingproduct 22, 384 mg, white foam, 65% yield and unreacted alkene 20, 63mg, 17%.

Preparation of N-Methylaniline 23:

To a solution of aniline 22 (178 mg, 0.219 mmol, 1.0 eq.) in THF (0.34mL) at 0° C. was added lithium hexamethyldisalazide (0.66 mL, 0.66 mmol,3.0 eq., lithium hexamethyldisalazide 1.0 M in THF). The resultingmixture was stirred at 0° C. during 60 min and added methyl iodide(0.085 mL, 1.35 mmol, 6.0 eq.). The reaction mixture was stirred 10 minat 0° C., then allowed to warm up to rt and stirred at that temperatureduring 21 h. A saturated solution of NH₄Cl and methyl-t-butyl ether wereadded. Layers were separated and the aqueous one was extracted twicewith methyl-t-butyl ether. The combined organic layers were washed withbrine, dried with sodium sulfate, filtered and concentrated to dryness.The resulting foam was purified by chromatography on silica gel 230-400Mesh using the following solvent gradient: 15% and 22% ethylacetate/hexane to elute. This procedure afforded N-methylaniline 23, 148mg, white foam, 82% yield.

Preparation of Macrocyclic Precursor 24:

To N-methylaniline 23 (119 mg, 0.143 mmol, 1.0 eq.) was added a solutionof TBAF (1.0 mL, 1.0 mmol, 7.0 eq.) buffered with imidazolehydrochloride. That buffered solution of TBAF was prepared by adding 5.3g of imidazole hydrochloride to 100 mL of TBAF 1.0 M in THF and bystirring till complete dissolution. The reaction mixture was stirred at65° C. during 16 h, cooled down to rt, added a saturated solution ofNH₄Cl and methyl-t-butyl ether. The layers were separated and theaqueous one was extracted twice with methyl-t-butyl ether. The combinedorganic layers were washed with brine, dried with sodium sulfate,filtered and concentrated to dryness. The resulting foam was purified bychromatography on silica gel 230-400 Mesh using the following solventgradient: 30% and 50% ethyl acetate/hexane to elute. This procedureafforded macrocylclic precursor 24, 94.0 mg, white foam, 92% yield.

Preparation of Macrolactone 25:

To a solution of macrocyclic precursor 24 (47.0 mg, 0.0658 mmol, 1.0eq.) in THF (6.6 mL) was added potassium hexamethyldisalazide 0.5 M intoluene (0.265 mL, 0.133 mmol, 2.0 eq.). Stirred 10 min at rt, quenchedwith a saturated solution of NH₄Cl to bring pH to neutral.Methyl-t-butyl ether was added, layers were separated, and the aqueousone was extracted with methyl-t-butyl ether. The combined organic layerswere washed with brine, dried with sodium sulfate, filtered andconcentrated to dryness. The resulting foam was purified bychromatography on silica gel 230-400 Mesh using the following solventgradient: 15% and 25% ethyl acetate/hexane to elute. This procedureafforded macrolactone 25, 23.5 mg, white foam, 55% yield.

Preparation of Ether 26:

To a solution of macrolactone 25 (17.9 mg, 0.0272 mmol, 1.0 eq.) indichloromethane (1.0 mL) at 0° C. was added N,N-diisopropylethylamine(0.125 ml, 0.712 mmol, 26.2 eq.) and MOMCl (0.042 mL, 0.550 mmol, 20.0eq.). The reaction mixture was allowed to warm up to rt and stirred atthat temperature during 3.75 h at which point it was quenched with asaturated solution of NH₄Cl. Methyl-t-butyl ether was added, layers wereseparated, and the aqueous one was extracted twice with methyl-t-butylether. The combined organic layers were washed with brine, dried withsodium sulfate, filtered and concentrated to dryness. The resulting foamwas purified by chromatography on silica gel 230-400 Mesh using thefollowing solvent gradient: 30% and 45% ethyl acetate/hexane to elute.This procedure afforded ether 26, 18.2 mg, white foam, 96% yield.

Preparation of Allylic Alcohol 27:

To a solution of ether 26 (18.2 mg, 0.026 mmol, 1.0 eq.) indichloromethane (0.60 mL) at rt was added water (0.12 mL) and DDQ (12.0mg, 0.0517 mmol, 2.0 eq.). The resulting mixture was stirred at rtduring 3.5 h at which point it was cooled down to 0° C. and quenchedwith a 1:1 solution of NaHCO₃ saturated and 10% Na₂S₂O₃ in water.Methyl-t-butyl ether was added, layers were separated, and the aqueousone was extracted twice with methyl-t-butyl ether. The combined organiclayers were washed with brine, dried with sodium sulfate, filtered andconcentrated to dryness. The resulting foam was purified by HPTLCprep-plates using 35% ethyl acetate/hexane to elute. This procedureafforded alcohol 27, 12.6 mg, white foam, 84% yield.

Note: Alcohol 27 is an intermediate of the first-generation synthesis ofER-806821, its conversion into ER-806821 can be achieved using theDess-Martin periodinane and the TFA procedures.

C) Synthesis of ER-807563

Preparation of N-Ethylaniline 39:

To a solution of aniline 22 (386 mg, 0.474 mmol, 1.0 eq.) in DMPU (1.6mL) at 0° C. was added lithium hexamethyldisalazide (0.950 mL, 0.950mmol, 2.0 eq., lithium hexamethyldisalazide 1.0 M in THF). The resultingmixture was stirred at 0° C. during 30 min and added ethyl iodide (0.230mL, 2.88 mmol, 6.1 eq.). The reaction mixture was stirred 10 min at 0°C., then allowed to warm up to rt and stirred at that temperature during1 h. A saturated solution of NH₄Cl, methyl-t-butyl ether and n-hexaneswere added. Layers were separated and the aqueous one was extractedtwice with methyl-t-butyl ether/hexane (1:1 mixture). The combinedorganic layers were washed with brine/water (1:1 mixture), then withbrine, dried with sodium sulfate, filtered and concentrated to dryness.The resulting foam was purified by chromatography on silica gel 230-400Mesh using the following solvent gradient: 15% and 20% ethylacetate/hexane to elute. This procedure afforded N-ethyl-aniline 39, 349mg, white foam, 87% yield.

Preparation of Macrocyclic Precursor 40:

N-ethylaniline 39 (817 mg, 0.970 mmol, 1.0 eq.) was added a solution ofTBAF (6.8 mL, 6.8 mmol, 7.0 eq.) previously buffered with imidazolehydrochloride. That buffered solution of TBAF was prepared by adding 5.3g of imidazole hydrochloride to 100 mL of TBAF 1.0 M in THF and bystirring till complete dissolution. The reaction mixture was stirred at65° C. during 16 h, cooled down to rt, added a saturated solution ofNH₄Cl, water and methyl-t-butyl ether. The layers were separated and theaqueous one was extracted twice with methyl-t-butyl ether. The combinedorganic layers were washed with brine/water (1:1 mixture), then washedwith brine, dried with sodium sulfate, filtered and concentrated todryness. The resulting foam was purified by chromatography on silica gel230-400 Mesh using the following solvent gradient: 20%, 30% and 45%ethyl acetate/hexane to elute. This procedure afforded macrocylclicprecursor 40, 635 mg, white foam, 90% yield.

Preparation of Macrolactone 41:

To a solution of macrocyclic precursor 40 (303 mg, 0.416 mmol, 1.0 eq.)in THF (40 mL) was added potassium hexamethyldisalazide (1.70 mL, 0.85mmol, 2.0 eq., potassium hexamethyldisalazide 0.5 M in toluene) over 3min. The reaction mixture was stirred 10 min at rt and quenched with asaturated solution of NH₄Cl. Methyl-t-butyl ether and water were added,layers were separated, and the aqueous one was extracted withmethyl-t-butyl ether. The combined organic layers were washed with asaturated solution of NH₄Cl, then washed with brine, dried with sodiumsulfate, filtered and concentrated to dryness. The resulting foam waspurified by chromatography on silica gel 230-400 Mesh using thefollowing solvent gradient: 15% and 18% ethyl acetate/hexane to elute.This procedure afforded macrolactone 41, 134 mg, white foam, 48% yield.

Preparation of Allylic Alcohol 42:

To a solution of macrolactone 41 (146.5 mg, 0.219 mmol, 1.0 eq.) indichloromethane (0.50 mL) was added water (0.50 mL). The resultingmixture was cooled down to 0° C. and was added a saturated solution ofDDQ in dichloromethane (3.0 mL, 0.265 mmol, 1.2 eq., this saturatedsolution of DDQ in dichloromethane contained at rt 20.5 mg of DDQ 98%pure/mL of dichloromethane). The biphasic reaction mixture was stirred 5min at 0° C., then 6 h at rt. At that point, it was cooled down to 0°C., added dichloromethane and quenched with a 1:1 solution of NaHCO₃saturated and 10% Na₂S₂O₃ in water. Stirring was continued for 15 min.Layers were separated and the aqueous one was extracted withdichloromethane. The combined organic layers were washed with a 1:1solution of NaHCO₃ saturated and 10% Na₂S₂O₃ in water, then washed withbrine, dried with sodium sulfate, filtered and concentrated to dryness.The resulting foam was purified by chromatography on silica gel 230-400Mesh using the following solvent gradient: dichloromethane, 3%, 5% and7.5% methyl-t-butyl ether/dichloromethane to elute. This procedureafforded allylic alcohol 42, 102 mg, white foam, 85% yield.

Preparation of Enone 43:

To a solution of allylic alcohol 42 (20.5 mg, 0.0372 mmol, 1.0 eq.) inn-heptane (1.0 mL) at rt was added MnO₂ (16 mg, 0.184 mmol, 5 eq.). Theresulting heterogeneous mixture was stirred at 65° C. during 12 h. Thisprocess gave only a small amount of desired material and in order topush the reaction to completion, fresh MnO₂ (16 mg) was added on a dailybasis until the reaction went to completion after one week of stirringat 65° C. Upon completion, ethyl acetate was added and the solids werefiltered off on a short silica gel column (230-400 Mesh) using ethylacetate as solvent. The filtrate was concentrated down to dryness andpurified by chromatography on silica gel 230-400 Mesh using 10%acetone/toluene as solvent. This procedure afforded enone 43, 7.0 mg,white foam, 35% yield (not optimized).

Preparation of ER-807563:

Enone 43 is treated according to the same TEA protocol as for thepreparation of ER-805940.

Preparation of NF2561

To a solution of 3 (10.0 g, 44.2 mmol) in DME (500 mL), pyridine (5.36mL, 66.3 mmol, 1.5 eq.) followed by nitronium tetrafluoroborate (8.8 g,66.3 mmol, 1.5 eq.) were added at −60° C. in dry ice-acetone bath andstirred for 1 h at −50° C. Then, pyridine (5.36 mL, 66.3 mmol, 1.5 eq.)followed by nitronium tetrafluoroborate (8.8 g, 66.3 mmol, 1.5 eq.) wereadded again at −60° C. in dry ice-acetone bath and stirred for 1 h at−50° C. One more time, pyridine (5.36 mL, 66.3 mmol, 1.5 eq.) followedby nitronium tetrafluoroborate (8.8 g, 66.3 mmol, 1.5 eq.) were added at−60 C in dry ice-acetone bath and stirred for 1 h at −50° C. Thereaction mixture was quenched at −58° C. with sat. NH₄Cl (1 L) andextracted with EtOAc (2×1 L). The organic layers were washed with waterand brine, dried over Na₂SO₄ and concentrated in vacuo. The crudeproduct was purified on silica gel column (Merck 230-400 mesh) withhexanes/EtOAc, 3:1 to give 9.59 g (35.4 mmol, 80%) of desired product asa yellow crystal.

To a mixture of 4 (10.55 g, 38.9 mmol) and pyridine (15.7 mL, 194 mmol,5 eq.) in CH₂Cl₂ (100 mL), Tf₂O (9.8 mL, 58.3 mmol, 1.5 eq.) wasgradually added at 0° C. during 30 min and the reaction mixture wasstirred for 45 min. The reaction mixture was quenched with sat. NH₄Cl(200 mL) and extracted with EtOAc (2×500 mL). The organic layers werewashed with water and brine, dried over Na₂SO₄ and concentrated invacuo. The crude product was purified on silica gel column (Merck230-400 mesh) with hexanes/EtOAc, 4:1 to give 13.9 g (34.5 mmol, 89%) ofdesired product as a yellow crystal.

A mixture of 5 (13.9 g, 34.5 mmol) and N-methylbenzylamine (22.7 mL,175.9 mmol, 5.1 eq.) in THF (175 mL) was refluxed overnight. Thereaction mixture was diluted with EtOAc (750 mL) and washed with water,10% KHSO₄, water and brine, dried over Na₂SO₄ and concentrated in vacuo.The crude product was purified on silica gel column (Merck 230-400 mesh)with hexanes/EtOAc, 20:1, 9:1 to give 9.7 g (25.9 mmol, 75%) of desiredproduct as a colorless oil.

A mixture of 6 (9.2 g, 24.6 mmol) and 20% Pd(OH)₂/C (1.75 g) in EtOH(200 mL) was hydrogenated under 1 atm of hydrogen for 4 hrs. Thecatalyst was filtered off and the filtrate was concentrated in vacuo.The crude diamine was obtained as yellow oil. It was used withoutpurification.

A mixture of the crude diamine, triethyl orthoformate (13.1 mL, 78.7mmol, 3.2 eq.) and montmorillonite KSF (2.7 g) in EtOH (200 mL) wasrefluxed for 100 min.

The insoluble material was filtered off and the filtrate wasconcentrated in vacuo. The crude product was purified on silica gelcolumn (Merck 230-400 mesh, 350 g) with hexanes/EtOAc, 1:1, 1:2 andEtOAc to give 84% (2 steps) of desired product as a yellow oil.

To a solution of 7 (5.75 g, 21.8 mmol) in CH₂Cl₂ (125 mL), TFA (16.8 mL,218 mmol, 10 eq.) was added at 0° C. Then, the reaction mixture waswarmed to room temperature and stirred for 1.5 hrs. The reaction mixturewas poured into ice water and neutralized with NaHCO₃ and extracted withEtOAc (800 mL+2×125 mL). The organic layers were washed with water andbrine, dried over Na₂SO₄. Na₂SO₄ was filtered and washed with CH₂Cl₂ andthe filtrate was concentrated in vacuo. The crude product was obtainedas white crystal and use without purification for the next step.

To a mixture of 60% NaH in mineral oil (2.3 g, 56.6 mmol, 2.6 eq.) andDMF (40 mL), a solution of the crude phenol 8 in DMF (60 mL) wasgradually added at 0° C. and stirred for 1 hr. Then, TBDPSCl (9.6 mL,37.0 mmol, 1.7 eq.) was added and the reaction mixture was warmed toroom temperature and stirred for 1.5 hr. The reaction mixture was pouredslowly in sat. NH₄Cl at 0° C. and extracted with EtOAc (3×150 mL). Theorganic layers were washed with water (×2) and brine, dried over Na₂SO₄and concentrated in vacuo. The crude product was purified on silica gelcolumn (Merck 230-400 mesh) with hexane/EtOAc, 3:1, 1:1, 2:3 to give8.89 g (19.4 mmol, 89% 2 steps) of desired product as a colorless oil.

A mixture of 9 (8.39 g, 18.3 mmol), NBS (3.6 g, 20.1 mmol, 1.1 eq.) andAIBN (1.2 g, 7.3 mmol, 0.4 eq.) in CCl₄ (250 mL) was heated slowly to50° C. and stirred for 4 hrs. The reaction mixture was cooled to roomtemperature and insoluble material was filtered off with celite, rinsedwith CCl₄ and the filtrate was concentrated in vacuo. The crude bromidewas used without purification. To a mixture of Cs₂CO₃ (9.2 g, 28.4 mmol,1.55 eq.) and DMF (90 mL), PhSH (2.9 mL, 28.4 mmol, 1.55 eq.) was addedat room temperature and the reaction mixture was stirred for 50 min.Then, a solution of the crude bromide in DMF (210 mL) was added and thereaction mixture was stirred for 1 hr. The reaction mixture was quenchedwith sat. NH₄Cl (300 mL) and extracted with EtOAc (3×300 mL). Theorganic layers were washed with water (×2) and brine, dried over Na₂SO₄and concentrated in vacuo. The crude product was purified on silica gelcolumn (Merck 230-400 mesh, 550 g) with hexanes/EtOAc, 2:1, 1:1, 1:3 togive 8.52 g (15.0 mmol, 82% 2 steps) of desired product.

To a solution of 10 (8.47 g, 14.9 mmol) in THF (200 mL), TBAF (1.0Msolution in THF, 22.4 mL, 1.5 eq.) was added at 0° C. and the reactionmixture was warmed to room temperature and stirred for 75 min. Thereaction mixture was quenched with sat. NH₄Cl (150 mL) and extractedwith EtOAc (3×200 mL). The organic layer was washed with water andbrine, dried over Na₂SO₄ and concentrated in vacuo. The crude productwas purified on silica gel column (Merck 230-400 mesh, 550 g) withhexanes/EtOAc, 2:1, 1:1, 1:2 to give 4.7 g (14.3 mmol, 96%) of desiredproduct.

To a solution of phenol 11 (4.41 g, 13.4 mmol) in DMF (100 mL) wereadded DBU (3.0 mL, 20.1 mmol, 1.5 eq.) and MOMCl (1.5 mL, 20.1 mmol, 1.5eq.) at rt. and the reaction mixture was stirred for 1.5 hr. Then, DBU(1.5 mL, 10.1 mmol, 0.75 eq.) and MOMCl (0.75 ml, 10.1 mmol, 0.75 eq.)were added and the reaction mixture was stirred for 1 hr.

The reaction mixture was quenched with sat. NH₄Cl (100 mL) and extractedwith EtOAc (3×150 mL). The organic layers were washed with water andbrine, dried over Na₂SO₄ and concentrated in vacuo. The crude productwas purified on silica gel column (Merck 230-400 mesh) withhexanes/EtOAc, 1:1, and 1:3 to give 87% of desired product

A mixture of 12 (4.5 g, 12.1 mmol), KOH 2M aq sol. (30 mL, 60.4 mmol, 5eq.) and DMSO (100 mL) was stirred at 80° C. for 1.5 hrs. The reactionmixture was cooled and adjusted to pH5 with 1N HCl (Be careful, MOMgroup can be easily hydrolyzed if pH too acidic). The resulted crystal(desired product) was filtered and washed water. Total 3.22 g of crudeproduct was obtained.

11) Esterification

To a solution of PPh₃ (2.3 eq) in THF (30 mL), DEAD (2.6 eq.) was addedat 0° C. and the reaction mixture was stirred for 30 min. Then, amixture of 13 (1 eq) and TMS ethanol (1.5 eq.) in THF (50 mL) was addedand the reaction mixture was warmed to room temperature and stirred forovernight. Furthermore, PPh₃ (2.3 eq.) and DEAD (2.6 eq.) was added andstirred for 10 min. Then, TMS ethanol (1.5 eq.) was added and stirredfor 20 min. The reaction mixture was concentrated in vacuo. Et₂O wasadded to the residue and stirred. The resulted solid (Ph₃P(O)) wasfiltered off and the filtrate was concentrated in vacuo. The crudeproduct was purified on silica gel column (Merck 230-400 mesh) withCH₂Cl₂/MeOH, 200:1, 100:1 to give a mixture of desired product and1,2-dicarbethoxyhydrazine (not separated). This mixture was purified onsilica gel column (Merck 230-400 mesh) with hexanes/EtOAc, 2:1, 1:1, and2:3 to give the desired product as white crystal.

To a solution of 14 (730 mg, 1.59 mmol, 1.5 eq.) and iodide 15 (639 mg,1.06 mmol, 1 eq.) in dry THF (25 mL)-HMPA (2.5 mL), LiHMDS (1.0Msolution in THF, 2.65 mL, 2.5 eq.) was added over a period of 25 min at−50° C. and the reaction mixture was for 30 min. Then, LiHMDS (1.0Msolution in THF, 1.3 mL, 1.25 eq.) was added at −50° C. and the reactionwas stirred for 1 hr. The reaction mixture was quenched with sat. NH₄Cl(50 mL) and extracted with EtOAc (3×50 mL). The organic layer was washedwith water and brine, dried over Na₂SO₄ and concentrated in vacuo. Thecrude product was purified on silica gel column (Merck 230-400 mesh)with hexanes/EtOAc, 4:1, 1:1, 1:3 to give 440 mg (0.471 mmol, 44%) ofdesired product as a colorless amorphous.

To a solution of 16 (980 mg, 1.22 mmol) in CH₂Cl₂ (45 mL), mCPBA (0.5 eqcalculated if mCPBA was 100%, 91 mg) was added at 0° C. and stirred for25 min, then mCPBA (91 mg) was added and stirred for 25 min, mCPBA (45mg) was added and stirred for 25 min, mCPBA (35 mg) was added andstirred for 20 min. The reaction mixture was quenched with sat. Na₂S₂O₃(80 mL) and extracted with EtOAc (400 ml+2×80 mL). The organic layer waswashed with sat. NaHCO₃ (×2) and brine, dried over Na₂SO₄ andconcentrated in vacuo to give 1.02 g of crude sulfoxide as colorlessamorphous.

A solution of crude sulfoxide and Et₃N (10 drops) in toluene (50 mL) wasrefluxed for 1 hr. The reaction mixture was concentrated in vacuo andthe crude product was purified on silica gel column (Merck 230-400 mesh)with hexanes/EtOAc, 3:1, 1:1, 1:3 to give 980 mg (1.19 mmol, 98% 2steps) of desired product as colorless amorphous.

To a solution of 18 (924 mg, 1.12 mmol) in THF (25 mL), Imidazole HCl(293 mg, 2.8 mmol, 2.5 eq) and TBAF (1.0M solution in THF, 5.6 ml, 5 eq)were added and the mixture was heated to 50° C. After 1 hr and 3 hrs,Imidazole HCl (293 mg, 2.8 mmol, 2.5 eq.) and TBAF (1.0M solution inTHF, 5.6 mL, 5 eq.) were added. After 24 hrs, TBAF (1.0M solution inTHF, 11.2 mL, 10 eq) was added and the reaction mixture was stirred for84 hrs at 50° C. The reaction mixture was quenched with sat. NH₄Cl (40mL) and saturated with NaCl (10 mL) and extracted with EtOAc (5×100 mL).The organic layers were dried over Na₂SO₄ and concentrated in vacuo. Thecrude product was purified on silica gel column (Merck 230-400 mesh)with EtOAc, EtOAc/MeOH, 99:1, 95:5 to give 740 mg of desired product(not pure) as a colorless amorphous.

To a refluxed mixture of 2-chloro-1-methylpyridinium iodide (4.21 mmol,4.0 eq.) and n-Bu₃N (1.0 mL, 4.21 mmol, 4 eq.) in CH₂Cl₂ (50 mL), asolution of 19 (640 mg, 1.05 mmol) in CH₂Cl₂ (25 mL) was added drop-wisewith syringe pump during 1 hr and then the reaction mixture was stirredfor 30 min. The reaction mixture was concentrated in vacuo and theresidue was diluted with EtOAc (750 mL), washed with water, sat. NH₄Cl,water and brine, dried over Na₂SO₄, filtered and concentrated in vacuo.The crude product was purified on silica gel column (Merck 230-400 mesh)with hexanes/EtOAc, 1:1, 1:5 CH₂Cl₂/MeOH, 95:5 to give 560 mg of desiredproduct (not pure) as colorless amorphous.

The hydrolysis was carried out as it described before in the synthesisof ER803064.

To a solution of 21 (260 mg, 0.596 mmol) in CH₂Cl₂ (45 mL), powderedMolecular Sieves 4 A (640 mg) and PCC (642 mg, 2.98 mmol, 5 eq.) wereadded at room temperature and the reaction mixture was stirred for 1hrs. The reaction mixture was filtered through Celite and the filtratewas concentrated in vacuo. The crude product was purified on silica gelcolumn (Merck 230-400 mesh) with CH₂Cl₂/MeOH, 99:1, 95:5 to give 185 mgof desired product (including pyridinium impurity).

To a solution of 22 (185 mg, 0.382 mmol) in CH₂Cl₂ (10 mL), TFA (0.88mL, 11.5 mmol, 30 eq.) was added at 0° C. and the reaction mixture waswarmed to room temperature and stirred for 1.5 hrs. The reaction mixturewas concentrated in vacuo. The crude product was purified on silica gelcolumn (Merck 230-400 mesh) with CH₂Cl₂/MeOH, 98:2, 95:5, 92:8 to give120 mg (0.300 mmol, 79%, 50% 2 steps) of desired product, NF2561. Thepure product was lyophilized with Water/MeCN, 1:1 to give a white foam.

ER805911 and ER805977:

These analogs were prepared using appropriate alternative reagents usingthe above synthesis. The modified steps are described below:

A mixture of the crude diamine (24.5 mmol), triethyl acetylformate (13.5mL, 3 eq.) and montmorillonite KSF (2.5 g) in EtOH (150 mL) was refluxedfor 2 h. The insoluble material was filtered off and the filtrate wasconcentrated in vacuo. The crude product was purified on silica gelcolumn (Merck 230-400 mesh, 350 g) with hexane/EtOAc, 1:1, 1:2 and EtOActo give 81% (2 steps) of desired product as yellow oil. The product wascarried forward in an analogous manner as described for NF2561 to giveER805911.

A mixture of triflate (9.3 g, 23 mmol) and N-ethylbenzylamine (15.2 mL,117.8 mmol, 5.1 eq.) in THF (175 mL) was refluxed 26 h. The reactionmixture was diluted with EtOAc (750 mL) and washed with water, 10%KHSO₄, water and brine, dried over Na₂SO₄ and concentrated in vacuo. Thecrude product was purified on silica gel column (Merck 230-400 mesh)with hexane/EtOAc, 20:1, 9:1 to give 6.25 g (16.1 mmol, 70%) of desiredproduct as a colorless oil. It was carried forward similarly asdescribed for NF2561 to give ER805977.

Preparation of C10 Analogs, ER804747:

The suspension of selenium dioxide (677 mg) and 531-YW-4 (1.96 g) indioxane (30 mL) was kept at 70° C. for 10 hours. The mixture wasconcentrated and purified with flush chromatograph (hexanes/acetate 5/1to 2/1) to give 593-YJ-22-1 (396 mg) and 593-YJ-22-2 (683 mg).

Sodium hydride (60%, 59 mg) was added to the solution of 593-YJ-22-2(788 mg) in DMF (15 mL), followed by the addition of methoxymethylchloride (160 mg) at room temperature. The mixture was kept stirring atroom temperature overnight and quenched with aqueous ammonium chloride.The aqueous phase was extracted with ether and the combined organicphase was concentrated. The residue was purified by flush chromatograph(hexane/acetate 4/1) to yield 593-YJ-25 (431 mg).

The solution of 593-YJ-25 (431 mg) and sodium hydroxide (1.0 N, 1.0 mL)in ethanol (5 mL) was kept stirring overnight at room temperature. Themixture was concentrated and diluted with aqueous ammonium chloride. Theaqueous phase was extracted with ether and the combined organic phasewas concentrated. The residue was purified by TLC (hexanes/acetate 2/1)to yield 593-YJ-29 (184 mg).

Dess-Martin periodinane was added to 593-YJ-29 (184 mg) in methylenechloride (10 mL) at room temperature. The mixture was diluted with etherin 2 hours and filtrated through Celite. The filtrate was concentratedand the residue was purified by TLC (hexanes/acetate 2/1) to give593-YJ-31 (138 mg).

n-Butyllithium (2.5 M, 0.56 mL) was added to 554-RB-228 (450 mg) in THF(5 mL) at −78° C. After one hour 593-YJ-31 (138 mg) was added. Thereaction was kept at 0° C. for one hour and warmed to room temperaturebefore it was quenched with aqueous ammonium chloride. The aqueous phasewas extracted with ether and the combined organic phase was dried oversodium sulfate. The solvent was stripped off and the residue waspurified with TLC (hexane/acetate 3/1) to 593-YJ-32 (168 mg, 75%).

The suspension of Rieke zink and 593-YJ-32 (168 mg) in methanol (10 mL)and water (1.0 mL) was kept at 70° C. for 4 hours. The mixture wasfiltrated through Celite and dried over sodium sulfate. The organics wasconcentrated and further dried azeotropically to give 593-YJ-33 (200mg). The crude 593-YJ-33 was subjected to triethylamine (2.7 mL) andbenzoyl chloride (1.1 mL) and purified with TLC (hexanes/acetate 4/1) togive 593-YJ-35 (358 mg).

The solution of 593-YJ-35 (358 mg) and imidazole hydrochlorite bufferedTBAF (1.0 M, 0.96 mL) in THF (10 mL) was kept stirring overnight at 50°C., and then diluted with water. The aqueous phase was extracted withether and concentrated. The residue was purified with TLC (methylenechloride/methanol, 10/1) to give 593-YJ-36 (41 mg).

593-YJ-36 (41 mg) was added to the reflux of 2-chloro-1-methylpyridiumiodide (52 mg) and tributylamine (43 mg) in methylene chloride (20 ml).After 2 hours reflux the mixture was stirred overnight. The mixture wasdiluted with ether and washed with HCl (1.0 N) and water. The residuewas purified with TLC (hexane/acetate 1/1) to give 593-YJ-39-1 (4.3 mg).

The solution of 593-YJ-39 (4.3 mg) and sodium hydroxide (1.0 N, 1.0 mL)in ethanol (5 mL) was kept stirring overnight at room temperature. Themixture was concentrated and diluted with aqueous ammonium chloride. Theaqueous phase was extracted with ether and the combined organic phasewas concentrated. The residue was purified by TLC (hexanes/acetate 2/1)to yield 593-YJ-57 (4.0 mg).

The suspension of MnO, (90%, 15 mg) and 593-YJ-57 (4.0 mg) in methylenechloride (2 mL) was kept stirring overnight. The mixture was filtratedthrough Celite and concentrated. The residue was purified with TLC(hexanes/acetate 2/3) to give 593-YJ-58 (2.0 mg).

Hydrofluoric acid (49%, 0.6 mL) was added to 593-YJ-58 (2.0 mg) inacetonitrile (1.5 mL) and stirred for 20 minutes. The mixture wasdiluted with water and extracted with methylene chloride. The organicphase was concentrated and purified with a short silica gel pad toproduce 593-YJ-59 (0.5 mg, ER-804747).

Preparation of C15-Methoxy-Analog, NF1872

To a solution of 3,4,5-trimethoxytoluene (5.47 g, 30 mmol) in DME (80mL), CuBr₂ (6.8 g, 30.45 mmol) was gradually added and the mixture wasstirred at room temperature for 30 min. Then, CuBr2 (8.4 g, 37.61 mmol)was added in several times. The mixture was stirred for 12 hrs. Theinsoluble material was filtered and the filtrate was concentrated. Thecrude product was purified on silica gel column with hexane/EtOAc, 4:1to give 7.5 g of NY-60.

NY-60 (7.39 g, 28.3 mmol) was dissolved in Et₂O (300 mL) and cooled to−78° C., under nitrogen. Then, n-BuLi (1.6M/hexane, 21 mL, 33.6 mmol)was slowly added and the reaction was stirred at −78° C. for 40 min. Dryice was added to the solution, then the solution was allowed to warm tort and was stirred for 15 min. The mixture was quenched with water,acidified with 2N HCl, extracted with EtOAc (×2). The organic layerswere washed with water, brine and dried over MgSO₄, filtered andconcentrated to give 6.38 g of NY-61.

To a solution of NY-61 (6.37 g, 28.16 mmol) in DMF (200 mL), Cs₂CO₃(9.17 g, 28.14 mmol) was added and stirred at room temperature for 10min. Then, MeI (10 mL, 160.6 mmol) was added and the reaction mixturewas stirred for 12 hrs. The reaction mixture was poured into ice-cooledsat. NH₄Cl and extracted with EtOAc (×3). The organic layers were washedwith water (×3), brine, dried over Na₂SO₄, filtered and concentrated.The crude product was purified on silica gel column with hexane/EtOAc,4:1 to give 6.07 g of NY-62.

NY-62 (6.07 g, 25.26 mmol) was dissolved in CH₂Cl₂ (80 mL) and cooled to−78° C., under nitrogen. Then, BCl₃ (1M/CH₂Cl₂, 26 mL, 26 mmol) wasslowly added and the reaction was stirred at −78° C. for 1 hr. Thesolution was stirred at 0° C. for 15 min, at room temperature for 1 hr.To the mixture which was recooled to −78° C., additional BCl₃(1M/CH₂Cl₂, 52 mL, 52 mmol) was slowly added and the solution wasallowed to warm to room temperature and was stirred for 15 hrs. Thereaction mixture was poured into ice-water and extracted with EtOAc(×2). The organic layers were washed with water (×2), brine, dried overMgSO₄, filtered and concentrated. The crude product was purified byrecrystallization from hexane/EtOAc to give 4.1 g of NY-63.

To a mixture of NY-63 (4.09 g, 19.27 mmol), MeOH (1.77 m L), ^(i)Pr₂NEt(3.7 mL, 21.24 mmol) and MeCN (80 mL), trimethylsilyldiazomethane(2M/hexane, 10.6 mL, 21.2 mmol) gradually added at 30° C. The reactionmixture was allowed to warm to room temperature and stirred for 24 hrs.The reaction mixture was quenched with water and diluted with EtOAc. Theorganic layer was washed with 10% citric acid and the aqueous layer wasreextracted with EtOAc. The organic layers were washed with water,brine, dried over Na₂SO₄, filtered and concentrated. The crude productwas purified on silica gel column with hexane/EtOAc, 20:1, 15:1, 10:1 togive 2.29 g of mixture of NY-64 and NY-65.

Using same procedure for NY-07, NY-65 was converted to NY-66 (1.73 g).(NY-64 was separated by column chromatography)

Using same procedure for 10, NY-66 (1.72 g, 6.41 mmol) was converted toNY-67 (2.07 g).

Using same procedure for NY-09, NY-67 (2.06 g, 5.47 mmol) was convertedto NY-68 (1.54 g).

Using same procedure for 509-HD-209, NY-68 (1.48 g, 4.43 mmol) wasconverted to NY-69 (1.62 g).

Using same procedure for 509-HD-212, NY-69 (1.62 g, 4.28 mmol) wasconverted to NY-70 (1.53 g).

Using same procedure for 509-HD-213, NY-70 (1.51 g, 4.14 mmol) wasconverted to NY-71 (1.08 g).

Using same procedure for 16, NY-27 (511 mg, 0.707 mmol) was converted toNY-72 (971 mg).

Using same procedure for 18, NY-72 (971 mg, 0.707 mmol) was converted toNY-33 (521 mg).

Using same procedure for 509-HD-116, NY-73 (521 mg, 0.549 mmol) wasconverted to NY-74 (444 mg). NY-74 was used without purification for thenext step.

Using same procedure for 509-HD-118, NY-74 (444 mg, 0.549 mmol)

was converted to NY-75 (86 mg) and NY-76 (114 mg).

Using same procedure for TM-13, NY-75 (45 mg, 0.094 mmol) was convertedto NY-77 (19.2 mg).

Using same procedure for NF-0675, NY-77 (18 mg, 0.038 mmol) wasconverted to NF-1872 (12.6 mg).

Preparation of C16 Analogs: NF0934, NF1418 and NF1419

Synthetic Procedure for NF-0934

Using same procedure for 554-RB-238, 531-yw-2-2 (5 g, 19.21 mmol) wasconverted to NY-20 (5.08 g). NY-20 was used without purification for thenext step.

To a suspension of lithium acetylide-ethylenediamine complex (24.92 g,0.271 mol) in DMSO (250 mL), (S)-propylene oxide (14.3 g, 0.246 mol) wasslowly added at 0° C. Then, the mixture was warmed to room temperatureand stirred for 24 hrs. The mixture was poured into ice-water andextracted with Et₂O (×4). The organic layers were washed with sat.NH₄Cl, brine and dried over MgSO₄, filtered and concentrated. The crudeproduct was used without purification for the next step.

Using same procedure for 554-RB-225, NY-21 (8 g, 95.1 mmol) wasconverted to NY-22 (27.33 g).

Using same procedure for NY-01, NY-20 (5.08 g, 19.21 mmol) was convertedto NY-23 (4.22 g) as one of diastereomers.

Using same procedure for 343-yw-279, NY-23 (4.2 g, 7.23 mmol) wasconverted to NY-24 (3.7 g).

To a solution of NY-24 (3.69 g, 6.33 mmol) in CH₂Cl₂ (70 mL),2,6-lutidine (3.7 mL, 31.8 mmol) and TBSOTf (3.63 mL, 15.8 mmol) wereadded at 0° C. Then, the mixture was warmed to room temperature andstirred for 30 min. The mixture was quenched with MeOH and poured intocold sat. NaHCO₃ and extracted with EtOAc. The organic layer was washedwith water, 5% citric acid, water, sat. NaHCO₃, brine and dried overNa₂SO₄, filtered and concentrated. The crude product was purified onsilica gel column with hexane/EtOAc, 100:1, 50:1, 30:1 to give 4.23 g ofNY-25.

NY-25 (4.2 g, 6.03 mmol) was dissolved in dry Et₂O (50 mL) and thesolution was cooled to 0° C. in ice/water bath. Then LiBH₄ (135 mg, 6.2mmol) was added portionwise, the mixture was allowed to warm slowly tort and stirred for 2 days after which a saturated solution of NH₄Cl wasadded slowly. The mixture was extracted with EtOAc and the organicextract was washed with a saturated solution of NH₄Cl, water, brine,dried with anhydrous Na₂SO₄, filtered and concentrated. The crudeproduct was purified on silica gel column with hexane/EtOAc, 8:1 to give3.55 g of NY-26.

Using same procedure for 554-RB-260, NY-26 (568 mg, 0.927 mmol) wasconverted to NY-27 (565 mg).

Using same procedure for 10, NY-28 (2.7 g, 15 mmol) was converted toNY-29 (2.45 g).

Using same procedure for 509-HD-212, NY-29 (2.45 g, 8.5 mmol) wasconverted to NY-11 (2.19 g).

Using same procedure for 509-HD-213, NY-30 (2.18 g, 3.06 mmol) wasconverted to NY-31 (2.35 g).

Using same procedure for 16, NY-27 (260 mg, 0.36 mmol) was converted toNY-32 (229 mg).

Using same procedure for 18, NY-32 (229 mg, 0.236 mmol) was converted toNY-33 (136 mg).

Using same procedure for 509-HD-116, NY-33 (136 mg, 0.585 mmol) wasconverted to NY-34 (119 mg).

Using same procedure for TM-12, NY-34 (116 mg, 0.285 mmol) was convertedto NY-35 (146 mg).

Using same procedure for TM-13, NY-35 (80 mg, 0.206 mmol) was convertedto NY-36 (55 mg).

Using same procedure for NF-0675, NY-36 (52 mg, 0.135 mmol) wasconverted to NF-0934 (29 mg).

Synthetic Procedure for NF-1418

Using same procedure for 10, NY-144 (7.11 g, 29.84 mmol) was convertedto NY-145 (7.32 g).

Using same procedure for NY-45, NY-145 (7.32 g, 21.13 mmol) wasconverted to NY-146 (4.46 g).

Using same procedure for NY-48, NY-146 (318 mg, 1 mmol) was converted tomixture of NY-147 and NY-148 (305 mg).

Using same procedure for 509-HD-213, NY-147 and NY-148 (303 mg, 0.912mmol) was converted to NY-149 (297 mg) and NY-150 (42 mg).

Using same procedure for 16, NY-53 (1.81 g, 4.808 mmol) was converted toNY-151 (2.06 g).

Using same procedure for 18, NY-151 (306 mg) was converted to NY-152(250 mg).

Using same procedure for 509-HD-116, NY-152 (230 mg, 0.233 mmol) wasconverted to NY-153 (197 mg).

Using same procedure for TM-12, NY-153 (136 mg, 0.233 mmol) wasconverted to NY-154 (30 mg).

A mixture of NY-154 (28 mg, 0.0494 mmol), 1N NaOH (125 μL, 0.125 mmol)and DME (0.5 mL) was stirred at room temperature for 18 hrs. Thereaction mixture was washed with Et₂O. The aqueous layer was neutralizedwith sat. NH₄Cl and extracted with EtOAc. The organic layers were washedwith brine, dried over Na₂SO₄, filtered and concentrated to give 23 mgof NY-155. NY-155 was used without purification for the next step.

A mixture of NY-155 (22 mg, 0.0398 mmol), diphenylphosphoryl azide (8.6μL, 0.0399 mmol), Et₃N (4 mg, 0.0395 mmol), ^(t)BuOH (0.3 mL) andtoluene (1.5 mL) was refluxed for 3 hrs. The reaction mixture wasdiluted with EtOAc and washed with 5% citric acid, water, sat. NaHCO₃,brine, dried over Na₂SO₄, filtered and concentrated. The crude productwas purified on silica gel column with hexane/EtOAc, 5:1, 3:1 to give 12mg of NY-156.

Using same procedure for 509-HD-188, NY-156 (11 mg, 0.0176 mmol) wasconverted to NY-157 (7.5 mg).

Using same procedure for TM-13, NY-157 (7.5 mg, 0.0149 mmol) wasconverted to NY-158 (5.3 mg).

Using same procedure for NF-0675, NY-158 (5.2 mg, 0.0104 mmol) wasconverted to NF-1418 (4.7 mg).

Synthetic Procedure for NF-1419

Using same procedure for B2538, NF-1418 (4 mg, 0.00937 mmol) wasconverted to NF-1419 (2.3 mg).

Measurement of Effect of Compounds on TNF-α and β-Actin PLAP (PlacentalAlkaline Phosphatase) Transcription.

NF-κB is a critical nuclear factor to regulate various genes involved inimmune and inflammatory responses. (see, Ghosh et al, Annu Rev Immunol.1998, 16, 225). It is well characterized that TNFα gene transcription isregulated by NF-th activation (see, Drouet et al. J. Immunol. 1991, 147,1694), therefore, the assay with TNFα-PLAP transcription was employed toevaluate the inhibitory effect of test compounds on NF-κB activation.

A TNFα-PLAP plasmid (TNFα-promoter+5′-UTR (1.4 kb)+PLAP+SV40polyA+PGK-neo, Goto et al. Mol. Pharmacol. 1996, 49, 860) wasconstructed with slight modification in which TNFα-3′-UTR (772 b.p.) wasinserted between PLAP and SV40 polyA (TNFα-promoter+5′-UTR (1.4kb)+PLAP+TNFα-3′-UTR+SV40 polyA+PGKneo). Then the THP-1-33 cells wereestablished by stably transfecting the modified TNFα-PLAP plasmid intoTHP-1 cells (human acute monocytic leukemia). In order to simultaneouslyevaluate non-specific effects of test compounds on transcription, B164cells were also established by stably transfecting β-actin-PLAP plasmidβ-actin-promoter (4.3 kb)+PLAP+SV40 polyA+PGKneo) into THP-1 cells.THP-1-33 cells (TNFα-PLAP) produce PLAP activity by the stimulation withLPS; on the other hand, B164 cells β-actin-PLAP) constantly produce PLAPactivity without stimuli.

THP-1-33 cells and B164 cells were maintained in RPMI1640 containing 10%heat-inactivated endotoxin-free fetal bovine serum (FBS) and G418 (1mg/ml). These cells were seeded at a density of 1.0×10⁴ cells/well onto96-well plate, and then were cultured in the presence or absence of testcompounds for 30 min, followed by stimulation with 100 ng/mL oflipopolysaccharide (E. coli 0127:B08 or 011:B4). After the cultivationfor 40-48 hrs, culture supernatant was harvested and alkalinephosphatase activity in the supernatant was measured.

Alkaline phosphatase activity was quantified with the use of achemiluminescent substrate,4-methoxy-4-(3-phosphatephenyl)spiro[1,2-dioxetane-3,2′-adamantane]. Toinactivate tissue-nonspecific alkaline phosphatase mainly derived fromFBS, samples were heated at 65° C. for 30 min before thechemiluminescent assay. Aliquots of 10 μL of culture supernatant weremixed with 50 μL of assay buffer (0.28 M Na₂CO₃—NaHCO₃, pH 10.0,containing 8 mM MgSO₄) in a 96-well Microlite™ plate (opaque), and then50 μL of chemiluminescent substrate was added and mixed. After 60 minincubation at room temperature, steady state chemiluminesce was measuredwith a microplate luminometer.

The PLAP activity of each sample was calculated as follows:TNFα-PLAP % of control=(A−B)×100/(C−B)β-actin-PLAP % of control=(A)×100/(C)

A: sample/chemiluminescence of the sample cultured with the test drug &stimulated with LPS

B: blank/chemiluminescence of unstimulated sample

C: control/chemiluminescence of the sample cultured with LPS

The IC50 value of each test compound was calculated from dose-inhibitoryresponse curve.

Drug Trade Approval Drug Name Approved Use Manufacturer/Distributor Datealitretinoin Panretin Topical treatment of cutaneous lesions in LigandPharmaceuticals Feb. 02, patients with AIDS-related Kaposi's sarcoma.1999 allopurinol Zyloprim Patients with leukemia, lymphoma and solidGlaxo Smith Kline May 17, tumor malignancies who are receiving cancer1996 therapy which causes elevations of serum and urinary uric acidlevels and who cannot tolerate oral therapy. altretamine Hexalen Singleagent palliative treatment of patients US Bioscience Dec. 26, withpersistent or recurrent ovarian cancer 1990 following first-line therapywith a cisplatin and/or alkylating agent based combination. amifostineEthyol To reduce the cumulative renal toxicity US Bioscience Dec. 08,associated with repeated administration of 1995 cisplatin in patientswith advanced ovarian cancer amifostine Ethyol Reduction of platinumtoxicity in non-small US Bioscience Mar. 15, cell lung cancer 1996amifostine Ethyol To reduce post-radiation xerostomia for head USBioscience Jun. 24, and neck cancer where the radiation port 1999includes a substantial portion of the parotid glands. anastrozoleArimidex for the adjuvant treatment of postmenopausal AstraZeneca Sep.05, women with hormone receptor positive early 2002 breast canceranastrozole Arimidex Treatment of advanced breast cancer in AstraZenecaDec. 27, postmenopausal women with disease Pharmaceuticals 1995progression following tamoxifen therapy. anastrozole Arimidex Forfirst-line treatment of postmenopausal AstraZeneca Sep. 01, women withhormone receptor positive or Pharmaceuticals 2000 hormone receptorunknown locally advanced or metastatic breast cancer. arsenic TrisenoxSecond line treatment of relapsed or refractory Cell Therapeutic Sep.25, trioxide APL following ATRA plus an anthracycline. 2000 bexaroteneTargretin For the treatment by oral capsule of cutaneous LigandPharmaceuticals Dec. 29, manifestations of cutaneous T-cell lymphoma1999 in patients who are refractory to at least one prior systemictherapy. bexarotene Targretin For the topical treatment of cutaneousLigand Pharmaceuticals Jun. 28, manifestations of cutaneous T-celllymphoma 2000 in patients who are refractory to at least one priorsystemic therapy. bleomycin Blenoxane Sclerosing agent for the treatmentof Bristol-Myers Squibb Feb. 20, malignant pleural effusion (MPE) and1996 prevention of recurrent pleural effusions. busulfan Busulfex Use incombination with cyclophoshamide as Orphan Medical, Inc Feb. 04,intravenous conditioning regimen prior to allogeneic 1999 hematopoieticprogenitor cell transplantation for chronic myelogenous leukemia.busulfan oral Myleran Chronic Myelogenous Leukemia-palliative GlaxoSmith Kline Jun. 26, therapy 1954 capecitabine Xeloda Initial therapy ofpatients with metastatic Roche Apr. 30, colorectal carcinoma whentreatment with 2001 fluoropyrimidine therapy alone is preferred.Combination chemotherapy has shown a survival benefit compared to5-FU/LV alone. A survival benefit over 5_FU/LV has not been demonstratedwith Xeloda monotherapy. capecitabine Xeloda Treatment in combinationwith docetaxel of Roche Sep. 07, patients with metastatic breast cancerafter 2001 failure of prior anthracycline containing chemotherapycapecitabine Xeloda Treatment of metastatic breast cancer resistantRoche Apr. 30, to both paclitaxel and an anthracycline 1998 containingchemotherapy regimen or resistant to paclitaxel and for whom furtheranthracycline therapy may be contraindicated, e.g., patients who havereceived cumulative doses of 400 mg/m2 of doxorubicin or doxorubicinequivalents carboplatin Paraplatin Palliative treatment of patients withovarian Bristol-Myers Squibb Mar. 03, carcinoma recurrent after priorchemotherapy, 1989 including patients who have been previously treatedwith cisplatin. carboplatin Paraplatin Initial chemotherapy of advancedovarian Bristol-Myers Squibb Jul. 05, carcinoma in combination withother 1991 approved chemotherapeutic agents. carmustine Gliadel For usein addition to surgery to prolong Guilford Pharmaceuticals Sep. 23, withWafer survival in patients with recurrent Inc. 1996 Polifeprosanglioblastoma multiforme who qualify for 20 Implant surgery. celecoxibCelebrex Reduction of polyp number in patients with Searle Dec. 23, therare genetic disorder of familial 1999 adenomatous polyposis.chlorambucil Leukeran Chronic Lymphocytic Leukemia-palliative GlaxoSmith Kline therapy cisplatin Platinol as a single agent for patientswith transitional Bristol-Myers Squibb Apr. 22, cell bladder cancerwhich is no longer 1993 amenable to local treatments such as surgeryand/or radiotherapy. cisplatin Platinol Metastatic testicular-inestablished Bristol-Myers Squibb Dec. 19, combination therapy with otherapproved 1978 chemotherapeutic agents in patients with metastatictesticular tumors whoc have already received appropriate surgical and/orradiotherapeutic procedures. An established combination therapy consistsof Platinol, Blenoxane and Velbam. cisplatin Platinol Metastatic ovariantumors - in established Bristol-Myers Squibb Dec. 19, combinationtherapy with other approved 1978 chemotherapeutic agents: Ovarian-inestablished combination therapy with other approved chemotherapeuticagents in patients with metastatic ovarian tumors who have alreadyreceived appropriate surgical and/or radiotherapeutic procedures. Anestablished combination consists of Platinol and Adriamycin. Platinol,as a single agent, is indicated as secondary therapy in patients withmetastatic ovarian tumors refractory to standard chemotherapy who havenot previously received Platinol therapy. cisplatin- IntraDose notapproved Matrix Pharmaceutical epinephrine (R), FocaCis gel (R)cladribine Leustatin, 2- Treatment of active hairy cell leukemia. R. W.Johnson Feb. 26, CdA Pharmaceutical Research 1993 Institute cytarabineDepoCyt Intrathecal therapy of lymphomatous Skye Pharmaceuticals Apr.01, liposomal meningitis 1999 daunorubicin DanuoXome First linecytotoxic therapy for advanced, HIV Nexstar, Inc. Apr. 08, liposomalrelated Kaposi's sarcoma. 1996 daunorubicin, DaunorubicinLeukemia/myelogenous/monocytic/erythroid Bedford Labs Jan. 30,daunomycin of adults/remission induction in acute 1998 lymphocyticleukemia of children and adults. daunorubicin, Cerubidine In combinationwith approved anticancer Wyeth Ayerst Res Mar. 11, daunomycin drugs forinduction of remission in adult ALL. 1987 dexrazoxane Zinecard reducingthe incidence and severity of Pharmacia Oct. 31, cardiomyopathyassociated with doxorubicin 2002 administration in women with metastaticbreast cancer who have received a cumulative doxorubicin dose of 300mg/m2 and who will continue to receive doxorubicin therapy to maintaintumor control. It is not recommended for use with the initiation ofdoxorubicin therapy. dexrazoxane Zinecard Prevention of cardiomyopathyassociated with Pharmacia & Upjohn May 26, doxorubicin administrationCompany 1995 docetaxel Taxotere Treatment of patients with locallyadvanced or Aventis Pharmaceutical May 14, metastatic breast cancer whohave progressed 1996 during anthracycline-based therapy or have relapsedduring anthracycline-based adjuvant therapy. docetaxel Taxotere Forlocally advanced or metastatic non-small Aventis Pharmaceutical Dec. 23,cell lung cancer after failure of prior platinum- 1999 basedchemotherapy. docetaxel Taxotere For the treatment of locally advancedor Aventis Pharmaceutical Jun. 22, metastatic breast cancer which hasprogressed 1998 during anthracycline-based treatment or relapsed duringanthracycline-based adjuvant therapy. doxorubicin Adriamycin Antibiotic,antitumor agent. Pharmacia & Upjohn Dec. 23, PFS Injection Company 1987doxorubicin Doxil Treatment of AIDS-related Kaposi's sarcoma SequusPharmaceuticals, Nov. 17, liposomal in patients with disease that hasprogressed on Inc. 1995 prior combination chemotherapy or in patientswho are intolerant to such therapy. doxorubicin Doxil Treatment ofmetastatic carcinoma of the Sequus Pharmaceuticals, Jun. 28, liposomalovary in patient with disease that is refractory Inc. 1999 to bothpaclitaxel and platinum based regimens Elliott's B Elliott's B Diluentfor the intrathecal administration of Orphan Medical, Inc Sep. 27,Solution Solution methotrexate sodium and cytarabine for the 1996prevention or treatment of meningeal leukemia or lymphocytic lymphoma.epirubicin Ellence A component of adjuvant therapy in patients Pharmacia& Upjohn Sep. 15, with evidence of axillary node tumor Company 1999involvement following resection of primary breast cancer. estramustineEmcyt palliation of prostate cancer Pharmacia & Upjohn Dec. 24, Company1981 etoposide Etopophos Management of refractory testicular tumors,Bristol-Myers Squibb May 17, phosphate in combination with otherapproved 1996 chemotherapeutic agents. etoposide Etopophos Management ofrefractory testicular tumors Bristol-Myers Squibb Feb. 27, phosphate andsmall cell lung cancer. 1998 etoposide Etopophos Management of smallcell lung cancer, first- Bristol-Myers Squibb May 17, phosphate line, incombination with other approved 1996 chemotherapeutic agents. etoposide,VePesid In combination with other approved Bristol-Myers Squibb Dec. 30,VP-16 chemotherapeutic agents as first line treatment 1986 in patientswith small cell lung cancer. etoposide, Vepesid Refractory testiculartumors-in combination Bristol-Myers Squibb Nov. 10, VP-16 therapy withother approved chemotherapeutic 1983 agents in patients with refractorytesticular tumors who have already received appropriate surgical,chemotherapeutic and radiotherapeutic therapy. etoposide, Vepesid Incombination with other approved Bristol-Myers Squibb Dec. 30, VP-16chemotherapeutic agents as first line treatment 1986 in patients withsmall cell lung cancer. exemestane Aromasin Treatment of advance breastcancer in Pharmacia & Upjohn Oct. 21, postmenopausal women whose diseasehas Company 1999 progressed following tamoxifen therapy. fludarabineFludara Palliative treatment of patients with B-cell Berlex LaboratoriesInc. Apr. 18, lymphocytic leukemia (CLL) who have not 1991 responded orhave progressed during treatment with at least one standard alkylatingagent containing regimen. fluorouracil, Adrucil prolong survival incombination with ICN Puerto Rico Apr. 25, 5-FU leucovorin 1962fulvestrant Faslodex the treatment of hormone receptor-positive IPR Apr.25, metastatic breast cancer in postmenopausal 2002 women with diseaseprogression following antiestrogen therapy gemcitabine Gemzar For use incombination with cisplatin for the Eli Lilly And Company Aug. 25,first-line treatment of patients with inoperable, 1998 locally advanced(Stage IIIA or IIIB) or metastatic (Stage IV) non-small cell lungcancer. gemcitabine Gemzar Treatment of patients with locally advancedEli Lilly And Company May 15, (nonresectable stage II or III) ormetastatic 1996 (stage IV) adenocarcinoma of the pancreas. Indicated forfirst-line treatment and for patients previously treated with a 5-fluorouracil-containing regimen. gemtuzumab- Mylotarg Treatment of CD33positive acute myeloid Wyeth Ayerst Res May 17, ozogamicin leukemia inpatients in first relapse who are 2000 60 years of age or older and whoare not considered candidates for cytotoxic chemotherapy. goserelinZoladex Palliative treatment of advanced breast cancer AstraZeneca Dec.18, acetate Implant in pre- and perimenopausal women. Pharmaceuticals1995 hydroxyurea Hydrea Decrease need for transfusions in sickle cellBristol-Myers Squibb Feb. 25, Capsules anemia 1998 idarubicin IdamycinFor use in combination with other approved Adria Laboratories Sep. 27,antileukemic drugs for the treatment of acute 1990 myeloid leukemia(AML) in adults. idarubicin Idamycin In combination with other approvedPharmacia & Upjohn Feb. 17, antileukemic drugs for the treatment ofacute Company 1997 non-lymphocytic leukemia in adults. ifosfamide IFEXThird line chemotherapy of germ cell Bristol-Myers Squibb Dec. 30,testicular cancer when used in combination 1988 with certain otherapproved antineoplastic agents. imatinib Gleevec metastatic orunresectable malignant Novartis Feb. 01, mesylate gastrointestinalstromal tumors 2002 imatinib Gleevec Initial therapy of chronicmyelogenous Novartis May 10, mesylate leukemia 2001 irinotecan CamptosarTreatment of patients with metastatic Pharmacia & Upjohn Jun. 14,carcinoma of the colon or rectum whose Company 1996 disease has recurredor progressed following 5-FU-based therapy. irinotecan Camptosar Forfirst line treatment n combination with 5- Pharmacia & Upjohn Apr. 20,FU/leucovorin of metastatic carcinoma of the Company 2000 colon orrectum. irinotecan Camptosar Follow up of treatment of metastaticPharmacia & Upjohn Oct. 22, carcinoma of the colon or rectum whoseCompany 1998 disease has recurred or progressed following 5-FU-basedtherapy. letrozole Femara Treatment of advanced breast cancer inNovartis Jul. 25, postmenopausal women. 1997 letrozole Femara First-linetreatment of postmenopausal women Novartis Jan. 10, with hormonereceptor positive or hormone 2001 receptor unknown locally advanced ormetastatic breast cancer. leucovorin Wellcovorin, Leucovorin calcium isindicated fro use in Immunex Corporation Jun. 20, Leucovorin combinationwith 5-fluorouracil to prolong 1952 survival in the palliative treatmentof patients with advanced colorectal cancer. leucovorin Leucovorin Incombination with fluorouracil to prolong Lederle Laboratories Dec. 12,survival in the palliative treatment of patients 1991 with advancedcolorectal cancer. levamisole Ergamisol Adjuvant treatment incombination with 5- Janssen Research Jun. 18, fluorouracil aftersurgical resection in patients Foundation 1990 with Dukes' Stage C coloncancer. liposomal DaunoXome “DaunoXome is indicated as a first lineVestar, Inc Jan. 28, daunorubicin cytotoxic therapy for advanced HIV-1994 associated Kaposi's Sarcoma. DaunoXome is not recommended inpatients with less than advnanced HIV-related Kaposi's Sarcoma.”melphalan, L- Alkeran Systemic administration for palliative Glaxo SmithKline Nov. 18, PAM treatment of patients with multiple myeloma 1992 forwhom oral therapy is not appropriate. mesna Mesnex Prevention ofifosfamide-induced Asta Medica Dec. 30, hemorrhagic cystitis 1988methotrexate Methotrexate osteosarcoma Lederle Laboratories Apr. 07,1988 methoxsalen Uvadex For the use of UVADEX with the UVAR TherakosFeb. 25, Photopheresis System in the palliative 1999 treatment of theskin manifestations of cutaneous T-cell lymphoma (CTCL) that isunresponsive to other forms of treatment. mitomycin C Mitozytrex therapyof disseminated adenocarcinoma of Supergen Nov. 14, the stomach orpancreas in proven 2002 combinations with other approvedchemotherapeutic agents and as palliative treatment when othermodalities have failed. mitoxantrone Novantrone For use in combinationwith corticosteroids as Immunex Corporation Nov. 13, initialchemotherapy for the treatment of 1996 patients with pain related toadvanced hormone-refractory prostate cancer. mitoxantrone Novantrone Foruse with other approved drugs in the Lederle Laboratories Dec. 23,initial therapy for acute nonlymphocytic 1987 leukemia (ANLL) in adults.paclitaxel Paxene treatment of advanced AIDS-related Kaposi's BakerNorton Dec. 24, sarcoma after failure of first line or Pharmaceuticals,Inc 1997 subsequent systemic chemotherapy paclitaxel Taxol Forfirst-line therapy for the treatment of Bristol-Myers Squibb Apr. 09,advanced carcinoma of the ovary in 1998 combination with cisplatin.paclitaxel Taxol Treatment of breast cancer after failure ofBristol-Myers Squibb Apr. 13, combination chemotherapy for metastatic1994 disease or relapse within 6 months of adjuvant chemotherapy. Priortherapy should have included an anthracycline unless clinicallycontraindicated. paclitaxel Taxol for use in combination with cisplatin,for the Bristol-Myers Squibb Jun. 30, first-line treatment of non-smallcell lung 1998 cancer in patients who are not candidates for potentiallycurative surgery and/or radiation therapy. paclitaxel Taxol For theadjuvant treatment of node-positive Bristol-Myers Squibb Oct. 25, breastcancer administered sequentially to 1999 standard doxorubicin-containingcombination therapy. paclitaxel Taxol First line ovarian cancer with 3hour infusion. Bristol-Myers Squibb Jun. 20, 2000 paclitaxel Taxol Newdosing regimen for patients who have Bristol-Myers Squibb Jun. 22,failed initial or subsequent chemotherapy for 1994 metastatic carcinomaof the ovary paclitaxel Taxol Treatment of patients with metastaticBristol-Myers Squibb Dec. 29, carcinoma of the ovary after failure offirst- 1992 line or subsequent chemotherapy. paclitaxel Taxol secondline therapy for AIDS related Kaposi's Bristol-Myers Squibb Aug. 04,sarcoma. 1997 pamidronate Aredia Treatment of osteolytic bone metastasesof Novartis Sep. 22, breast cancer in conjunction with standard 1998antineoplastic therapy. Pegademase Adagen Enzyme replacement therapy forpatients with Enzon Mar. 21, (Pegademase severe combinedimmunodeficiency asa result 1990 Bovine) of adenosine deaminasedeficiency. pentostatin Nipent Single agent treatment for adult patientswith Parke-Davis Oct. 11, alpha interferon refractory hairy cellPharmaceutical Co. 1991 leukemia. pentostatin Nipent Single-agenttreatment for untreated hairy cell Parke-Davis Sep. 29, leukemiapatients with active disease as Pharmaceutical Co. 1993 defined byclinically significant anemia, neutropenia, thrombocytopenia, ordisease- related symptoms. (Supplement for front-line therapy.) porfimerPhotofrin For use in photodynamic therapy (PDT) for QLTPhototherapeutics Inc. Dec. 27, sodium palliation of patients withcompletely 1995 obstructing esophageal cancer, or patients withpartially obstructing esophageal cancer who cannot be satisfactorilytreated with ND- YAG laser therapy. porfimer Photofrin For use inphotodynamic therapy (PDT) for QLT Phototherapeutics Inc. Dec. 22,sodium reduction of obstruction and palliation of 1998 symptoms inpatients with completely or partially obstructing endobroncial nonsmallcell lung cancer (NSCLC). porfimer Photofrin For use in photodynamictherapy for treatment QLT Phototherapeutics Inc. Jan. 09, sodium ofmicroinvasive endobronchial nonsmall cell 1998 lung cancer in patientsfor whom surgery and radiotherapy are not indicated. streptozocinZanosar Antineoplastic agent. Pharmacia & Upjohn May 07, Company 1982talc Sclerosol For the prevention of the recurrence of Bryan Dec. 24,malignant pleural effusion in symptomatic 1997 patients. tamoxifenNolvadex For use in women with axillary node-negative AstraZeneca Jun.21, breast cancer adjuvant therapy. Pharmaceuticals 1990 tamoxifenNolvadex As a single agent to delay breast cancer AstraZeneca Dec. 03,recurrence following total mastectomy and Pharmaceuticals 1986 axillarydissection in postmenopausal women with breast cancer (T1-3, N1, M0)tamoxifen Nolvadex to reduce the incidence of breast cancer inAstraZeneca Oct. 29, women at high risk for breast cancerPharmaceuticals 1998 tamoxifen Nolvadex Metastatic breast cancer in men.AstraZeneca Apr. 01, Pharmaceuticals 1993 tamoxifen Nolvadex to reducethe incidence of breast cancer in AstraZeneca Oct. 29, women at highrisk for breast cancer Pharmaceuticals 1998 tamoxifen Nolvadex In womenwith DCIS, following breast AstraZeneca Jun. 29, surgery and radiation,Nolvadex is indicated Pharmaceuticals 2000 to reduce the risk ofinvasive breast cancer. tamoxifen Nolvadex Equal bioavailability of a 20mg Nolvadex AstraZeneca Mar. 21, tablet taken once a day to a 10 mgNolvadex Pharmaceuticals 1994 tablet taken twice a day. tamoxifenNolvadex For use in premenopausal women with AstraZeneca Mar. 16,metastatic breast cancer as an alternative to Pharmaceuticals 1989oophorectomy or ovarian irradiation temozolamide Temodar Treatment ofadult patients with refractory Schering Aug. 11, anaplastic astrocytoma,i.e., patients at first 1999 relapse with disease progression on anitrosourea and procarbazine containing regimen teniposide, Vumon Incombination with other approved Bristol-Myers Squibb Jul. 14, VM-26anticancer agents for induction therapy in 1992 patients with refractorychildhood acute lymphoblastic leukemia (all). topotecan HycamtinTreatment of patients with metastatic Glaxo Smith Kline May 28,carcinoma of the ovary after failure of initial 1996 or subsequentchemotherapy. topotecan Hycamtin Treatment of small cell lung cancersensitive Glaxo Smith Kline Nov. 30, disease after failure of first-line1998 chemotherapy. In clinical studies submitted to support approval,sensitive disease was defined as disease responding to chemotherapy butsubsequently progressing at least 60 days (in the phase 3 study) or atleast 90 days (in the phase 2 studies) after chemotherapy toremifeneFareston Treatment of advanced breast cancer in Orion Corp. May 29,postmenopausal women. 1997 tretinoin, Vesanoid Induction of remission inpatients with acute Roche Nov. 22, ATRA promyelocytic leukemia (APL) whoare 1995 refractory to or unable to tolerate anthracycline basedcytotoxic chemotherapeutic regimens. valrubicin Valstar For intravesicaltherapy of BCG-refractory Anthra --> Medeva Sep. 25, carcinoma in situ(CIS) of the urinary bladder 1998 in patients for whom immediatecystectomy would be associated with unacceptable morbidity or mortality.vinorelbine Navelbine Single agent or in combination with cisplatinGlaxo Smith Kline Dec. 23, for the first-line treatment of ambulatory1994 patients with unresectable, advanced non- small cell lung cancer(NSCLC). vinorelbine Navelbine Navelbine is indicated as a single agentor in GlaxoSmithKline Nov. 05, combination with cisplatin for thefirst-line 2002 treatment of ambulatory patients with unreseactable,advanced non-small cell lung cancer (NSCLC). In patients with Stage IVNSCLC, Navelbine is indicated as a single agent or in combination withcisplatin. In Stage III NSCLC, Navelbine is indicated in combinationwith cisplatin. zoledronate Zometa the treatment of patients withmultiple Novartis Feb. 22, myeloma and patients with documented bone2002 metastases from solid tumors, in conjunction with standardantineoplastic therapy. Prostate cancer should have progressed aftertreatment with at least one hormonal therapy

1. A method for treating rheumatoid arthritis, psoriasis, asthma,sepsis, inflammatory bowel disease, atopic dermatitis and Crohn'sdisease comprising: administering to a subject in need thereof atherapeutically effective amount of a compound

or pharmaceutically acceptable salt, ester, or salt of ester thereof;wherein R₁ is hydrogen, aliphatic, heteroaliphatic, alicyclic, or aryl;R₂ is methyl and R₃ is hydrogen or halogen; R₄ is hydrogen or halogen;R₅ is hydrogen or an oxygen protecting group; R₆ is hydrogen, hydroxyl,or protected hydroxyl; n is 1; R₇ is hydrogen; R₈ is hydrogen, halogen,hydroxyl, protected hydroxyl or alkyloxy; R₉ is NR₁₂R₁₃; wherein R₁₂ andR₁₃ are, independently for each occurrence, hydrogen, aliphatic,heteroaliphatic, alicyclic, or aryl; or a protecting group, and each ofR₁₂ and R₁₃ are optionally further substituted with one or moreoccurrences of hydroxyl, protected hydroxyl, alkyloxy, amino, protectedamino, alkylamino, aminoalkyl, or halogen, R₁₀ is hydroxyl, protectedhydroxyl, or amino; R₁₁ is hydrogen; X is O, NH, or N-alkyl; Y is CHR₁₇,O, CR₁₇ or NR₁₇; and Z is CHR₁₈, C═O, or CR₁₈, wherein each occurrenceof R₁₇ and R₁₈ is hydrogen, or R₁₇ and R₁₈ taken together is —O—, or—CH₂—, and Y and Z are connected by a single or double bond; andoptionally a pharmaceutically acceptable carrier or diluent.
 2. Themethod of claim 1, wherein the method is for treating rheumatoidarthritis.
 3. The method of claim 1, wherein the method is for treatingpsoriasis.
 4. The method of claim 1, wherein the method is for treatingasthma.
 5. The method of claim 1, wherein: wherein R₁ is hydrogen,straight or branched C₁₋₆alkyl, straight or branched C₁₋₆heteroalkyl, oraryl, wherein the alkyl, heteroalkyl, and aryl groups may optionally besubstituted with one or more occurrences of halogen, hydroxyl orprotected hydroxyl; R₂ is methyl and R₃ is hydrogen or halogen; R₄ ishydrogen or halogen; R₅ is hydrogen or an oxygen protecting group; R₆ ishydrogen, hydroxyl, or protected hydroxyl; R₇ is hydrogen; R₈ ishydrogen; R₉ is NR₁₂R₁₃; wherein R₁₂ and R₁₃ are, independently for eachoccurrence, hydrogen, aliphatic, heteroaliphatic, alicyclic, or aryl, ora protecting group, and each of R₁₂ and R₁₃ are optionally furthersubstituted with one or more occurrences of hydroxyl, protectedhydroxyl, alkyloxy, amino, protected amino, alkylamino, aminoalkyl, orhalogen; R₁₀ is hydroxyl, protected hydroxyl, or amino; R₁₁ is hydrogen;X is O, NH, or N-alkyl; Y is CHR₁₇, O, CR₁₇ or NR₁₇; and Z is CHR₁₈,C═O, or CR₁₈, wherein each occurrence of R₁₇ and R₁₈ is hydrogen, or R₁₇and R₁₈ taken together is —O—, or —CH₂—, and Y and Z are connected by asingle or double bond.
 6. The method of claim 5, wherein in the compoundX is oxygen.
 7. The method of claim 5, wherein in the compound R₄ ishalogen.
 8. The method of claim 5, wherein in the compound R₄ ishydrogen.
 9. The method of claim 5, wherein in the compound Y and Ztogether represent —CH═CH—.
 10. The method of claim 5, wherein in thecompound Y and Z together represent trans —CH═CH—.
 11. The method ofclaim 5, comprising administering a compound wherein R₁ and R₂ are eachmethyl and R₃ is hydrogen and the compound is of the structure:

wherein R₄-R₁₁, X, Y and Z are as defined in claim
 5. 12. The method ofclaim 11, wherein in the compound X is oxygen.
 13. The method of claim11, wherein in the compound R₄ is halogen.
 14. The method of claim 11,wherein in the compound R₄ is hydrogen.
 15. The method of claim 11,wherein in the compound Y and Z together represent —CH═CH—.
 16. Themethod of claim 11, wherein in the compound R₄ is hydrogen, and Y and Ztogether represent —CH═CH—.
 17. The method of claim 15 or 16, wherein inthe compound —CH═CH— is trans.
 18. The method of claim 5, wherein X isoxygen, R₁ and R₂ are each methyl, R₃ is hydrogen, R₄ is hydrogen and Yand Z together represent —CH═CH—.
 19. A method of claim 1, comprisingadministering a compound of the structure:

or pharmaceutically acceptable salt, ester, or salt of ester thereof.20. A method of claim 1, comprising administering a compound of thestructure:


21. A method of claim 1, comprising administering a compound of thestructure:

or pharmaceutically acceptable salt, ester, or salt of ester thereof.22. A method of claim 1, comprising administering a compound of thestructure:


23. A method of claim 1, comprising administering a compound of thestructure:

or pharmaceutically acceptable salt, ester, or salt of ester thereof.24. A method of claim 1, comprising administering a compound of thestructure:

or pharmaceutically acceptable salt, ester, or salt of ester thereof.25. A method of claim 1, comprising administering a compound of thestructure:

or pharmaceutically acceptable salt, ester, or salt of ester thereof.26. A method of claim 1, comprising administering a compound of the

or pharmaceutically acceptable salt, ester, or salt of ester thereof.27. A method for the treatment of psoriasis, comprising administering acompound according to any one of claims 5, 11, 19, 20, 21, and
 22. 28.The method of claim 27, wherein in the step of administering, thecomposition is administered topically.
 29. A method for the treatment ofpsoriasis, comprising administering a compound of the structure:

or pharmaceutically acceptable salt, ester, or salt of ester thereof.30. A method for the treatment of psoriasis, comprising administering acompound of the structure:


31. A method for the treatment of psoriasis, comprising administering acompound of the structure:

or pharmaceutically acceptable salt, ester, or salt of ester thereof.32. A method for the treatment of psoriasis, comprising administering acompound of the structure: